Product Citations: 145

Fig.2.D showing Western Blotting from the publication: Isoxazole-based molecules restore NK cell immune surveillance in hepatocarcinogenesis by targeting TM4SF5 and SLAMF7 linkage.
Fig.3.A showing Western Blotting from the publication: STING mediates nuclear PD-L1 targeting-induced senescence in cancer cells.
Fig.6.A showing Western Blotting from the publication: Centrosome function is critical during terminal erythroid differentiation.
Fig.7.A showing Western Blotting from the publication: Centrosome function is critical during terminal erythroid differentiation.
Fig.3.D showing Western Blotting from the publication: HMGB1 orchestrates STING-mediated senescence via TRIM30α modulation in cancer cells.
Fig.3.D showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
Fig.2.B showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
Fig.1.C showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
Fig.1.A showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
Fig.1.E showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
Fig.3.B showing Western Blotting from the publication: CDK8/19 inhibition induces premature G1/S transition and ATR-dependent cell death in prostate cancer cells.
Fig.7.B showing Western Blotting in a Homo sapiens (Human) sample from the publication: The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.
Fig.5.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.
Fig.1.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.
Fig.3.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.
Fig.2.A showing Western Blotting from the publication: Age-dependent defective TGF-beta1 signaling in patients undergoing coronary artery bypass grafting.
Fig.7.A showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
Fig.5.A showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
Fig.4.E showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
Fig.4.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
Fig.5.B showing Western Blotting in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
Fig.5.D showing Western Blotting in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
Fig.4.C showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
Fig.4.B showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
Fig.5.C showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Pten dose dictates cancer progression in the prostate.
25 images found

Distinct outcomes from targeted perturbations of the multi-subunit SCFSkp2 E3 ubiquitin ligase in blocking Trp53/Rb1-null prostate tumorigenesis.

In Communications Biology on 22 February 2025 by Xue, Y., Zhu, L., et al.

Identifying effective therapies targeting multi-protein complexes that lack catalytic sites or cofactor pockets remains a long-standing challenge. The proto-oncogene, ubiquitin E3 ligase SCFSkp2, is one such target. SCFSkp2 promotes the proteasomal degradation of the cyclin-dependent kinase inhibitor p27, which controls cell cycle progression. Targeted knockout of Rb1/Trp53 causes metastatic prostate cancer in mice; additional knockout of Skp2 completely blocks tumorigenesis. We compared gene-edited mice that carried two different single amino acid changes in the SCFSkp2 complex, structurally predicted to inhibit the degradation of p27. Mutation of the SCFSkp2 accessory protein Cks1 (Cks1N45R) completely blocked Rb1/Trp53-driven prostate tumorigenesis, phenocopying Skp2 knockout, whereas a mutation directly stabilizing p27 (p27T187A) did not. This was consistent with structural models that predicted the binding of both p27 and p27T187A to the SCFSkp2/Cks1/Cdk2/CyclinA/p27 complex, and their subsequent ubiquitination and degradation, albeit at different rates. Two binding modes, which differ in their dependence on phosphorylated T187, are predicted by the model. Studies confirmed the role of p27 in mediating tumorigenesis in Rb1/Trp53 mutant tumors and revealed a mutually destabilizing Skp2 and p27 feedback loop. The integration of gene editing, drug-surrogate mutations, and mouse tumor models offers a blueprint for studying SCFSkp2 and other multi-subunit biomedical targets.
© 2025. The Author(s).

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Isoxazole-based molecules restore NK cell immune surveillance in hepatocarcinogenesis by targeting TM4SF5 and SLAMF7 linkage.

In Signal Transduction and Targeted Therapy on 20 January 2025 by Kim, J. E., Kim, H. S., et al.

Dynamic communication between hepatocytes and the environment is critical in hepatocellular carcinoma (HCC) development. Clinical immunotherapy against HCC is currently unsatisfactory and needs more systemic considerations, including the identification of new biomarkers and immune checkpoints. Transmembrane 4 L six family member 5 (TM4SF5) is known to promote HCC, but it remains unclear how cancerous hepatocytes avoid immune surveillance and whether avoidance can be blocked. We investigated how TM4SF5-mediated hepatic tumorigenesis avoids surveillance by natural killer (NK) cells, which are prevalent in the liver, and whether the avoidance can be blocked by anti-TM4SF5 agents. We used comprehensive structure activity relationship analysis to identify TM4SF5-specific isoxazole (TSI)-based small molecules that inhibit TM4SF5-mediated effects. TM4SF5 expressed by hepatocytes reduced NK cell cytotoxicity by downregulating stimulatory ligands/receptors, including signaling lymphocytic activation molecule family member 7 (SLAMF7). TM4SF5 bound SLAMF7 depending on N-glycosylation and caused intracellular trafficking of SLAMF7 from the plasma membrane to lysosomes for degradation. TSI treatments in cell lines and animal models of HCC blocked this binding, intracellular trafficking, and downregulation, resulting in higher levels of stimulatory NK cell ligands. In mouse xenograft models, TSI treatment abrogated HCC development by increasing the abundance and dispersion of Slamf7-positive cells in liver tissues, recapitulating the phenotype of Tm4sf5-knockout mice and indicating TSI-mediated restoration of NK cell surveillance. These findings suggest that TSIs can inhibit TM4SF5-mediated liver carcinogenesis by increasing NK cell surveillance.
© 2025. The Author(s).

  • WB
  • Immunology and Microbiology
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Effects of phospholipase D1-inhibitory peptide on the growth and metastasis of gastric cancer cells.

In Molecules and Cells on 1 November 2024 by Kim, D., Yoon, M. S., et al.

Phospholipase D1 (PLD1) contributes to cancer development and progression through its effects on cell proliferation, survival, invasion, metastasis, angiogenesis, drug resistance, and modulation of the tumor microenvironment. Its central role in these processes makes it a promising target for novel cancer treatments aimed at inhibiting its activity and disrupting the signaling pathways it regulates. In this study, we aimed to investigate the effect of PLD1 inhibition on gastric cancer cell growth using a novel peptide inhibitor, TAT-TVTSP. PLD1, which plays a role in cancer progression, catalyzes the conversion of phosphatidylcholine into choline and phosphatidic acid through hydrolysis. To effectively target PLD1 in cells, we engineered TAT-TVTSP by fusing a PLD1-inhibitory peptide (TVTSP) with a cell-penetrating peptide (TAT). We observed that TAT-TVTSP effectively inhibited PLD1 activity in AGS gastric cancer cells. Moreover, TAT-TVTSP significantly inhibited the mammalian target of the rapamycin signaling pathway, including the phosphorylation of key downstream targets such as S6K1, AKT, S473, glycogen synthase kinase-3b, and forkhead box O1. TAT-TVTSP did not induce cell death, but it triggered cell cycle arrest by activating p21 and p27 via AKT phosphorylation. Functional assays revealed that TAT-TVTSP significantly impaired the colony-forming ability of AGS cells, thus inhibiting cell proliferation. Transwell and wound-healing assays revealed that this peptide disrupted the cellular behaviors critical to cancer progression, such as migration and invasion. In vivo, TAT-TVTSP significantly reduced tumor growth in the xenograft model of gastric cancer without any toxicity. Overall, our results suggest that TAT-TVTSP is a novel therapeutic agent for PLD1-mediated cancers.
Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.

  • Cancer Research
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PPTC7 antagonizes mitophagy by promoting BNIP3 and NIX degradation via SCFFBXL4.

In EMBO Reports on 1 August 2024 by Nguyen, G. D. T., Townsend, B., et al.

Mitophagy must be carefully regulated to ensure that cells maintain appropriate numbers of functional mitochondria. The SCFFBXL4 ubiquitin ligase complex suppresses mitophagy by controlling the degradation of BNIP3 and NIX mitophagy receptors, and FBXL4 mutations result in mitochondrial disease as a consequence of elevated mitophagy. Here, we reveal that the mitochondrial phosphatase PPTC7 is an essential cofactor for SCFFBXL4-mediated destruction of BNIP3 and NIX, suppressing both steady-state and induced mitophagy. Disruption of the phosphatase activity of PPTC7 does not influence BNIP3 and NIX turnover. Rather, a pool of PPTC7 on the mitochondrial outer membrane acts as an adaptor linking BNIP3 and NIX to FBXL4, facilitating the turnover of these mitophagy receptors. PPTC7 accumulates on the outer mitochondrial membrane in response to mitophagy induction or the absence of FBXL4, suggesting a homoeostatic feedback mechanism that attenuates high levels of mitophagy. We mapped critical residues required for PPTC7-BNIP3/NIX and PPTC7-FBXL4 interactions and their disruption interferes with both BNIP3/NIX degradation and mitophagy suppression. Collectively, these findings delineate a complex regulatory mechanism that restricts BNIP3/NIX-induced mitophagy.
© 2024. The Author(s).

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Targeted inhibition of SCFSKP2 confers anti-tumor activities resulting in a survival benefit in osteosarcoma.

In Oncogene on 1 March 2024 by Wang, J., Ferrena, A., et al.

Osteosarcoma(OS) is a highly aggressive bone cancer for which treatment has remained essentially unchanged for decades. Although OS is characterized by extensive genomic heterogeneity and instability, RB1 and TP53 have been shown to be the most commonly inactivated tumor suppressors in OS. We previously generated a mouse model with a double knockout (DKO) of Rb1 and Trp53 within cells of the osteoblastic lineage, which largely recapitulates human OS with nearly complete penetrance. SKP2 is a repression target of pRb and serves as a substrate recruiting subunit of the SCFSKP2 complex. In addition, SKP2 plays a central role in regulating the cell cycle by ubiquitinating and promoting the degradation of p27. We previously reported the DKOAA transgenic model, which harbored a knock-in mutation in p27 that impaired its binding to SKP2. Here, we generated a novel p53-Rb1-SKP2 triple-knockout model (TKO) to examine SKP2 function and its potential as a therapeutic target in OS. First, we observed that OS tumorigenesis was significantly delayed in TKO mice and their overall survival was markedly improved. In addition, the loss of SKP2 also promoted an apoptotic microenvironment and reduced the stemness of DKO tumors. Furthermore, we found that small-molecule inhibitors of SKP2 exhibited anti-tumor activities in vivo and in OS organoids as well as synergistic effects when combined with a standard chemotherapeutic agent. Taken together, our results suggest that SKP2 inhibitors may reduce the stemness plasticity of OS and should be leveraged as next-generation adjuvants in this cancer.
© 2024. The Author(s).

  • Cancer Research
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Isoxazole-based molecules restore NK cell immune surveillance in hepatocarcinogenesis by targeting TM4SF5 and SLAMF7 linkage.

In Signal Transduct Target Ther on 20 January 2025 by Kim, J. E., Kim, H. S., et al.

Fig.2.D
Fig.2.D showing Western Blotting from the publication: Isoxazole-based molecules restore NK cell immune surveillance in hepatocarcinogenesis by targeting TM4SF5 and SLAMF7 linkage.
  • WB
Isoxazoles abrogated HCC development via inhibitions of signal activity depending on TM4SF5 expression. a Scheme for TM4SF5-expressing SNU449T7 cell xenograft models with NOD-SCID male mice (6-week-old, n = 10). b Tumor volumes were measured twice a week with caliper and calculated. Tumor weight ... more
Isoxazoles abrogated HCC development via inhibitions of signal activity depending on TM4SF5 expression. a Scheme for TM4SF5-expressing SNU449T7 cell xenograft models with NOD-SCID male mice (6-week-old, n = 10). b Tumor volumes were measured twice a week with caliper and calculated. Tumor weight was measured right after sacrifice and tumor extraction. The maximal and minimal tumor weight values were eliminated as outliers for animal variances upon drug treatment, resulting in data values at n = 8 for graphs and tumor images. c Liver tissues were processed to immunostaining for the indicated molecules. Representative images were shown. Scale bars, 100 μm. d Subconfluent TM4SF5-negative SNU449Cp (Cp) or TM4SF5-positive SNU449T7 cells were treated with vehicle or isoxazoles (3 μM) for 24 h, prior to harvests and immunoblots. * in the immunoblots indicate non-specific bands. *P < 0.05, unpaired Student’s t tests or Two-way ANOVA. Data are represented as mean ± SEM. Data stands for three independent experiments. See also Fig. S4 less

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STING mediates nuclear PD-L1 targeting-induced senescence in cancer cells.

In Cell Death Dis on 15 September 2022 by Lee, J. J., Kim, S. Y., et al.

Fig.3.A
Fig.3.A showing Western Blotting from the publication: STING mediates nuclear PD-L1 targeting-induced senescence in cancer cells.
  • WB
STING mediates PD-L1 targeting-induced senescence.A–C B16-F10 cells were transfected with 100 nM Control (siC) or PD-L1 (siPD-L1) siRNA prior to treatment with 100 ng/mL Dox. Western blot analysis of the indicated proteins was performed on day 3 after Dox treatment (A). Endogenous STING and PD-L1... more
STING mediates PD-L1 targeting-induced senescence.A–C B16-F10 cells were transfected with 100 nM Control (siC) or PD-L1 (siPD-L1) siRNA prior to treatment with 100 ng/mL Dox. Western blot analysis of the indicated proteins was performed on day 3 after Dox treatment (A). Endogenous STING and PD-L1 expression were determined by confocal microscopy following staining with anti-STING or anti-PD-L1 antibodies with the appropriate secondary antibodies on day 3 after Dox treatment (B, C). Scale bars = 20 μm. D–G B16-F10 cells were individually transfected with 100 nM siRNAs against STING (siSTING), Control (siC), or PD-L1 (siPD-L1). Then, cells were treated with 100 ng/mL Dox and the morphological changes (D), relative cell numbers (E), and SA-β-Gal-positive cells (F, G) were analyzed on day 3 post treatment. β-actin was detected as a loading control. Scale bars = 20 μm (B, C) and 50 μm (D, F). The significance of the statistical difference among the three group was calculated using a one-way analysis of variance and Newman–Keuls. Quantitative data are expressed as the mean ± SD. **p < 0.01, ***p < 0.001. less

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Centrosome function is critical during terminal erythroid differentiation.

In EMBO J on 18 July 2022 by Tátrai, P. & Gergely, F.

Fig.6.A
Fig.6.A showing Western Blotting from the publication: Centrosome function is critical during terminal erythroid differentiation.
  • WB
Terminal erythroid differentiation is impaired in Trp53 and Cdk5rap2 double mutants both ex vivo and in vivoImmunoblot showing levels of TP53, P27, and P21 ex vivo cultured erythroid progenitors upon CB treatment. GAPDH was used as loading control.Quantification of mean protein levels from (A). N... more
Terminal erythroid differentiation is impaired in Trp53 and Cdk5rap2 double mutants both ex vivo and in vivoImmunoblot showing levels of TP53, P27, and P21 ex vivo cultured erythroid progenitors upon CB treatment. GAPDH was used as loading control.Quantification of mean protein levels from (A). Numbers in brackets represent number of embryos analyzed.Quantification of cell cycle profiles of ex vivo cultured Trp53HET or Trp53null TER119pos cells following CB treatment from Fig 7C. Numbers in brackets represent number of embryos analyzed.Quantification of number of cell divisions at 48 h (T48) of ex vivo culture following CB treatment. Ex vivo cultured Trp53HET or Trp53null EBs were labeled with PKH26 to measure cell divisions. Number of embryos analyzed is shown in brackets.Quantification of number of cell divisions of CB‐treated enucleating EBs with indicated DNA content from (D). Percentage of enucleating EBs in each category is shown below X‐axis. Number of embryos analyzed is shown in brackets.Quantification of cell cycle profiles of ex vivo cultured TER119pos cells from Fig 7E. Number of embryos analyzed is shown in brackets.Quantification of ex vivo cultured apoptotic (AnnexinVpos) TER119pos cells with indicated genotypes after 48 h (T48). Number of embryos analyzed is shown in brackets.Complete blood count analysis from adult mice with genotypes as indicated. The number of mice analyzed is shown in brackets. RBC = red blood cell. MCV = mean corpuscular volume.Data information: Box plots show 5th and 95th (whiskers) and 25th, 50th, and 75th percentiles (boxes). Statistical analysis was based on the number of embryos (B‐G) or number of mice (H). All statistical significances were determined by one‐way ANOVA with Tukey's multiple comparisons test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. less

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Centrosome function is critical during terminal erythroid differentiation.

In EMBO J on 18 July 2022 by Tátrai, P. & Gergely, F.

Fig.7.A
Fig.7.A showing Western Blotting from the publication: Centrosome function is critical during terminal erythroid differentiation.
  • WB
Elevated TP53 activity resulting from centrosome or CDK5RAP2 loss is not responsible for abnormal erythroblast differentiationImmunoblot analysis of TP53, P27, and P21 levels in ex vivo cultured Cdk5rap2 wild‐type (WT), heterozygous (HET), and null EBs after 24 (T24) and 48 (T48) hours. GAPDH was... more
Elevated TP53 activity resulting from centrosome or CDK5RAP2 loss is not responsible for abnormal erythroblast differentiationImmunoblot analysis of TP53, P27, and P21 levels in ex vivo cultured Cdk5rap2 wild‐type (WT), heterozygous (HET), and null EBs after 24 (T24) and 48 (T48) hours. GAPDH was used as loading control. ** Indicates unspecific band.Quantification of mean protein levels from (A). Numbers in brackets represent number of embryos analyzed.Quantification of cell cycle profiles of ex vivo cultured Trp53HET or Trp53null enucleating EBs with or without CB treatment. EBs were pulsed with BrdU according to Fig 3A. Numbers in brackets represent number of embryos analyzed. See Fig EV6C for cell cycle profiles of TER119pos cells.Quantification of ex vivo cultured Trp53HET or Trp53null enucleating EBs and reticulocytes after 48 h (T48) upon CB treatment. Numbers in brackets represent number of embryos analyzed.Quantification of cell cycle profiles of ex vivo cultured enucleating EBs with the indicated genotypes after 48 h (T48). Number of embryos analyzed is shown in brackets. See Fig EV6F for cell cycle profiles of TER119pos cells.Quantification of ex vivo cultured enucleating EBs and reticulocytes with the indicated genotypes after 48 h (T48). Number of embryos analyzed is shown in brackets.Model shows the origin of EBs enucleating with 2N versus 4N DNA content and the consequences on reticulocyte production.Data information: Box plots show 5th and 95th (whiskers) and 25th, 50th, and 75th percentiles (boxes). All statistical analysis was based on number of embryos. All statistical significances were determined by one‐way ANOVA with Tukey's multiple comparisons test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. less

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HMGB1 orchestrates STING-mediated senescence via TRIM30α modulation in cancer cells.

In Cell Death Discov on 8 February 2021 by Lee, J. J., Park, I. H., et al.

Fig.3.D
Fig.3.D showing Western Blotting from the publication: HMGB1 orchestrates STING-mediated senescence via TRIM30α modulation in cancer cells.
  • WB
TRIM30α targeting induced senescence via STING.A–C B16 cells were transfected with Si C or Si TRIM30α 24 h prior to Dox treatment for 3 days. Cells were photographed (A), and SA-β-GAL-positive cells (B) were counted (C). B16 Cells were transfected with various concentrations of Si TRIM30α, and we... more
TRIM30α targeting induced senescence via STING.A–C B16 cells were transfected with Si C or Si TRIM30α 24 h prior to Dox treatment for 3 days. Cells were photographed (A), and SA-β-GAL-positive cells (B) were counted (C). B16 Cells were transfected with various concentrations of Si TRIM30α, and western blotting was performed on day 3 after Dox treatment (D). E, F Tumors were generated by implantation of B16-F10 cells into mice. On day 14 after implantation, tumors were exposed to IR. Three hours later, tumors were harvested and analyzed by western blotting. G–K TRIM30α WT and KO mouse BMDMs were employed for analyzing the effects of TRIM30 on senescence. Real-time PCR of TRIM30α was performed with TRIM30α-WT and -KO BMDMs (G). TRIM30α-WT and -KO BMDMs were treated with 100 ng/mL Dox, and morphological changes (H: left, enlarged images: right), images (I), and counts (J) of SA-β-GAL-positive cells were evaluated on day 3 after treatment. Western blotting was performed at the indicated times after treatment with 100 ng/mL Dox (K). less

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p27Kip1 regulates alpha-synuclein expression.

In Oncotarget on 27 March 2018 by Gallastegui, E., Domuro, C., et al.

Fig.3.D
Fig.3.D showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
  • WB
E2F4 and p130 regulate α-SYN expression(A) Control MEFS and E2F4 KO MEFs were assessed for E2F4 and α-SYN by WB. Tubulin was used as a loading control. (B) α-SYN mRNA levels were determined by qPCR in the same cells as in (A). Results are expressed relative to Control 1 (Ctrl1). (C) Control MEFS ... more
E2F4 and p130 regulate α-SYN expression(A) Control MEFS and E2F4 KO MEFs were assessed for E2F4 and α-SYN by WB. Tubulin was used as a loading control. (B) α-SYN mRNA levels were determined by qPCR in the same cells as in (A). Results are expressed relative to Control 1 (Ctrl1). (C) Control MEFS (WT) and p130 KO MEFs were assessed for p130 and α-SYN by WB. Tubulin was used as a loading control. (D) Control MEFS (WT) and double KO MEFs p130/p27 (DKO) were assessed for p130, p27 and α-SYN by WB. Tubulin was used as a loading control. In all experiments results are the mean value ± SD of at least three independent experiments. Statistical analyses were performed using the Student’s t-test. **P<0.01 and ***P<0.001. less

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p27Kip1 regulates alpha-synuclein expression.

In Oncotarget on 27 March 2018 by Gallastegui, E., Domuro, C., et al.

Fig.2.B
Fig.2.B showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
  • WB
p21 regulates α-SYN expression(A) Levels of α-SYN and p21 were determined in c17.2 control cells infected with ShRNA control or ShRNA for p21. Tubulin was used as a loading control. (B) Cells from the p27KO clone P1 were infected with ShRNA control or ShRNA for p21 and subsequently assessed for p... more
p21 regulates α-SYN expression(A) Levels of α-SYN and p21 were determined in c17.2 control cells infected with ShRNA control or ShRNA for p21. Tubulin was used as a loading control. (B) Cells from the p27KO clone P1 were infected with ShRNA control or ShRNA for p21 and subsequently assessed for p27, p21 and α-SYN protein by WB. Tubulin was used as a loading control. (C) α-SYN mRNA levels were determined by qPCR in the same cells as in (B). (D) α-SYN protein levels were determined in c17.2 cells treated or not with the Cdk inhibitor Roscovitine (20 μM). In all experiments results are the mean value ± SD of at least three independent experiments. Statistical analyses were performed using the Student’s t-test. ***P<0.001. less

Collected and cropped from Oncotarget by CiteAb, provided under a CC-BY license

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p27Kip1 regulates alpha-synuclein expression.

In Oncotarget on 27 March 2018 by Gallastegui, E., Domuro, C., et al.

Fig.1.C
Fig.1.C showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
  • WB
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. Aft... more
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. After normalizing data respect tubulin, a correlation between p27 and p21 levels was analyzed using the regression lineal program SPSS. The significance was P<0.05. The value of the Pearson correlation was -0,762. (C) The levels of p27 and α-SYN were determined by WB in two control clones (1 and 2) and two p27KO clones (P1 and P2). Tubulin was used as a loading control. (D) Levels of α-SYN mRNA were determined in the same samples by qPCR. Results are expressed relative to Control 1 (Ctrl1). (E) Cells from the p27KO clone P1 were infected with lentiviruses with an empty vector (Ф) or with wild-type p27 (p27wt). Then, protein levels of p27 and α-SYN protein were determined by WB. Tubulin was used as a loading control. (F) α-SYN mRNA levels were determined by qPCR in the same cells as in (E). In all experiments results are the mean value ± SD of at least three independent experiments. Statistical analyses were performed using the Student’s t-test. ***P<0.001. less

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p27Kip1 regulates alpha-synuclein expression.

In Oncotarget on 27 March 2018 by Gallastegui, E., Domuro, C., et al.

Fig.1.A
Fig.1.A showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
  • WB
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. Aft... more
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. After normalizing data respect tubulin, a correlation between p27 and p21 levels was analyzed using the regression lineal program SPSS. The significance was P<0.05. The value of the Pearson correlation was -0,762. (C) The levels of p27 and α-SYN were determined by WB in two control clones (1 and 2) and two p27KO clones (P1 and P2). Tubulin was used as a loading control. (D) Levels of α-SYN mRNA were determined in the same samples by qPCR. Results are expressed relative to Control 1 (Ctrl1). (E) Cells from the p27KO clone P1 were infected with lentiviruses with an empty vector (Ф) or with wild-type p27 (p27wt). Then, protein levels of p27 and α-SYN protein were determined by WB. Tubulin was used as a loading control. (F) α-SYN mRNA levels were determined by qPCR in the same cells as in (E). In all experiments results are the mean value ± SD of at least three independent experiments. Statistical analyses were performed using the Student’s t-test. ***P<0.001. less

Collected and cropped from Oncotarget by CiteAb, provided under a CC-BY license

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p27Kip1 regulates alpha-synuclein expression.

In Oncotarget on 27 March 2018 by Gallastegui, E., Domuro, C., et al.

Fig.1.E
Fig.1.E showing Western Blotting from the publication: p27Kip1 regulates alpha-synuclein expression.
  • WB
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. Aft... more
p27 regulates α-SYN expression(A) c17.2 cells were knocked out for p27 by using the CRISPR/Cas9 methodology. Then, severalcontrol c17.2 clones were assessed for p27 and p21 by WB. Tubulin was used as a loading control. (B) The bands from Figure 1A were quantified by using the image J program. After normalizing data respect tubulin, a correlation between p27 and p21 levels was analyzed using the regression lineal program SPSS. The significance was P<0.05. The value of the Pearson correlation was -0,762. (C) The levels of p27 and α-SYN were determined by WB in two control clones (1 and 2) and two p27KO clones (P1 and P2). Tubulin was used as a loading control. (D) Levels of α-SYN mRNA were determined in the same samples by qPCR. Results are expressed relative to Control 1 (Ctrl1). (E) Cells from the p27KO clone P1 were infected with lentiviruses with an empty vector (Ф) or with wild-type p27 (p27wt). Then, protein levels of p27 and α-SYN protein were determined by WB. Tubulin was used as a loading control. (F) α-SYN mRNA levels were determined by qPCR in the same cells as in (E). In all experiments results are the mean value ± SD of at least three independent experiments. Statistical analyses were performed using the Student’s t-test. ***P<0.001. less

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CDK8/19 inhibition induces premature G1/S transition and ATR-dependent cell death in prostate cancer cells.

In Oncotarget on 2 March 2018 by Nakamura, A., Nakata, D., et al.

Fig.3.B
Fig.3.B showing Western Blotting from the publication: CDK8/19 inhibition induces premature G1/S transition and ATR-dependent cell death in prostate cancer cells.
  • WB
Induction of DNA damage response followed by ATR-dependent cell death mediated by CDK8/19 inhibition(A) VCaP cells were treated with 30 nmol/L T-474 for 1–7 days. Cell cycle profile was analyzed using a flow cytometer. Line graphs show the population of cells in G1, S, and sub-G1 phase (N = 3, me... more
Induction of DNA damage response followed by ATR-dependent cell death mediated by CDK8/19 inhibition(A) VCaP cells were treated with 30 nmol/L T-474 for 1–7 days. Cell cycle profile was analyzed using a flow cytometer. Line graphs show the population of cells in G1, S, and sub-G1 phase (N = 3, mean with SD). (B) VCaP cells were treated with 30 nmol/L T-474 or 300 nmol/L T-418 for 1–3 days. Cell lysates were analyzed by western blot. (C) VCaP cells were treated with T-474 together with the ATR inhibitor VE-821, ATM inhibitor KU-55933, or DNA-PK inhibitor NU7026 as indicated for 4 days. (D) VCaP cells were treated with T-474 together with Z-VAD as indicated for 4 days. Cell viability was measured (N = 3, mean with SD). (E) VCaP cells were treated with T-474 together with ATRi (VE-821) or Z-VAD as indicated for 4 days. Caspase 3/7 activity was measured (N = 3, mean with SD). *P < 0.01 and **P < 0.000001. (F) VCaP cells were treated with T-474 together with SN-38, ETP (etoposide), or DOX (doxorubicin) as indicated for 4 days. Cell viability was measured (N = 3, mean with SD). N.S., not significant (P > 0.05). less

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The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.

In Sci Rep on 16 May 2016 by Ferrán, B., Martí-Pàmies, I., et al.

Fig.7.B
Fig.7.B showing Western Blotting in a Homo sapiens (Human) sample from the publication: The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.
  • WB
  • Homo sapiens (Human)
SMPX silencing does not affect HSMM differentiation.(a) Microphotographs from HSMM transfected with a siRNA against SMPX (siSMPX) or a control random siRNA (siRd) and then exposed to differentiation medium during five days. (b) SMPX mRNA levels (upper panel) and protein levels of SMPX, MyHC and p... more
SMPX silencing does not affect HSMM differentiation.(a) Microphotographs from HSMM transfected with a siRNA against SMPX (siSMPX) or a control random siRNA (siRd) and then exposed to differentiation medium during five days. (b) SMPX mRNA levels (upper panel) and protein levels of SMPX, MyHC and p27 (lower panel) in HSMM treated as indicated in (a) (n = 6). *P < 0.0001 vs. siRd. Levels of β-actin were used as a loading control. (c) Immunocytochemistry for MyHC and phalloidin-stained F-actin in cells treated as indicated in (a). Bars: 25 μm. less

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The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.

In Sci Rep on 16 May 2016 by Ferrán, B., Martí-Pàmies, I., et al.

Fig.5.C
Fig.5.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: The nuclear receptor NOR-1 regulates the small muscle protein, X-linked (SMPX) and myotube differentiation.
  • WB
  • Homo sapiens (Human)
NOR-1 knockdown prevents HSMM differentiation and down-regulates SMPX expression.HSMM were transfected with a siRNA against NOR-1 (siNOR-1) or a control random siRNA (siRd), and then were exposed to differentiation conditions during five days. (a) Microphographs (left) and immunofluorescence micr... more
NOR-1 knockdown prevents HSMM differentiation and down-regulates SMPX expression.HSMM were transfected with a siRNA against NOR-1 (siNOR-1) or a control random siRNA (siRd), and then were exposed to differentiation conditions during five days. (a) Microphographs (left) and immunofluorescence microscopy analysis showing filamentous actin (F-actin stained with phalloidin; right) from HSMM treated as indicated above. (b) NOR-1 and SMPX mRNA levels in cells transfected with siNOR-1 (black bars) or with siRd (white bars) (n = 6). *P < 0.0001 vs. siRd. (c) Representative Western blot showing protein levels of NOR-1, SMPX, and cell markers associated to HSMM differentiation (MyHC and p27) in HSMM treated as indicated above. Levels of proteins in non-transfected HSMM, both differentiated (D) and non-differentiated (ND), are also shown. Levels of β-actin were used as a loading control (n = 4). (d) Immunofluorescence microscopy analysis showing NOR-1 (green), SMPX (green) and MyHC (green) in cells as indicated in (a). Nuclear staining using Hoechst (blue) is also shown. Bars: 25 μm. less

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Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.

In Oncotarget on 20 February 2015 by Vosper, J., Masuccio, A., et al.

Fig.1.A
Fig.1.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.
  • WB
  • Homo sapiens (Human)
Lovastatin induces downregulation of the Skp2 protein(A) HeLa cells growing asynchronously or arrested in S-Phase by treatment with 2mM thymidine for 24 hours, were then treated or left untreated with 40μM lovastatin for a further 24 hours and analysed by (A) immunoblotting. (B) LnCAP and HeLa ce... more
Lovastatin induces downregulation of the Skp2 protein(A) HeLa cells growing asynchronously or arrested in S-Phase by treatment with 2mM thymidine for 24 hours, were then treated or left untreated with 40μM lovastatin for a further 24 hours and analysed by (A) immunoblotting. (B) LnCAP and HeLa cells (C) were incubated with 10μM and 40μM lovastatin respectively for the indicated times and analysed by immunoblotting. (D) HeLa cells stably expressing Skp2 from a PGK promoter were generated by lentiviral transfection. These cells were treated with 40μM lovastatin for 24 hours and then subjected to immunoblotting analysis. Percentages of cells in the different cell cycle phases were determined by flow cytometric analysis of three independent experiments using propidium iodine and BrdU double staining (Bar graph; data are represented as mean +/− SEM). (E) Control MEFs and p27 −/− MEFs were treated with 20μM lovastatin for 24 hours as indicated, harvested and analysed by western blotting analysis (left panel). Flow cytometry analysis of BrdU labeled cells to determine cell cycle phase distribution was performed. The percentage of cells in G1, S or G2/M phase is shown for at least three independent experiments (right panel). (F) Skp2 mRNA analysis of HeLa cells treated as described in Fig. 1A. Northern blots from three independent experiments were quantified using a PhosphorImager and the relative abundance of the Skp2 mRNA is shown (bar graph; data are represented as mean +/− SEM). less

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Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.

In Oncotarget on 20 February 2015 by Vosper, J., Masuccio, A., et al.

Fig.3.A
Fig.3.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Statin-induced depletion of geranylgeranyl pyrophosphate inhibits cell proliferation by a novel pathway of Skp2 degradation.
  • WB
  • Homo sapiens (Human)
Skp2 decline is due to blockade of geranylgeranylation(A) HeLa cells were treated with 40μM lovastatin in the presence of geranylgeranylpyrophosphate (GGPP) or farnesylpyrophosphate (FPP). After 24 hours incubation, cells were harvested and subjected to western blotting analysis. (B) HeLa cells w... more
Skp2 decline is due to blockade of geranylgeranylation(A) HeLa cells were treated with 40μM lovastatin in the presence of geranylgeranylpyrophosphate (GGPP) or farnesylpyrophosphate (FPP). After 24 hours incubation, cells were harvested and subjected to western blotting analysis. (B) HeLa cells were treated 20μM lovastatin, geranylgeranyltransferase inhibitor GGTI-298 or farnesyltransferase inhibitor FTI-277 for 36 hours and analysed by western blotting. (C) HeLa cells were treated with 10μM GGTI-298 for different times (as indicated) after which Skp2, p27 levels were determined by immunoblot analysis. (D) HeLa cells were treated with 10μM GGTI-298 in the presence and absence of 20mM MG132. After 24 hours, samples were harvested and analysed by western blotting. (E) Control MEFs and p27 −/− MEFs were treated with 10 μM GGTI-298 for 24 hours as indicated, harvested and analysed by western blotting analysis (left panel). Flow cytometry analysis of BrdU labeled cells to determine cell cycle phase distribution was performed. The percentage of cells in G1, S or G2/M phase is shown for at least three independent experiments (right panel). (F) Control MEFs and Cdh1−/− MEFs were treated with 10μM GGTI-298 for 24 hours then harvested and subjected to western blotting analysis. (G) Hela cells were treated with 50ng/ml Nocodazole for 18 hours after which they were supplemented with GGTI-298 at a final concentration of 10μM or solvent control. After a further 3 hours cells were supplemented with MG132 (20μM) or solvent and incubated for a further 9 hours. Cells were then harvested and processed for western blotting analysis. less

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Age-dependent defective TGF-beta1 signaling in patients undergoing coronary artery bypass grafting.

In J Cardiothorac Surg on 4 February 2014 by Redondo, S., Navarro-Dorado, J., et al.

Fig.2.A
Fig.2.A showing Western Blotting from the publication: Age-dependent defective TGF-beta1 signaling in patients undergoing coronary artery bypass grafting.
  • WB
Age-dependent decreased p27 in IMA from CABG patients. Panel A: Representative Western blot of p27 expression versus ERK in IMA from CABG patients. Panel B: p27 expression and age show a significant inverse correlation. Panel C: age-dependent p27 decrease remains statistically significant when co... more
Age-dependent decreased p27 in IMA from CABG patients. Panel A: Representative Western blot of p27 expression versus ERK in IMA from CABG patients. Panel B: p27 expression and age show a significant inverse correlation. Panel C: age-dependent p27 decrease remains statistically significant when compared among several age groups. *p < 0.05. less

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Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.

In PLoS Biol on 1 December 2013 by Lecona, E., Rojas, L. A., et al.

Fig.7.A
Fig.7.A showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
  • WB
SCML2B stabilizes p21.(A) Western blot analysis of p21, p27, and actin, using whole cell extracts from U2OS cells treated with two different siRNAs (#1, #2) for SCML2 or a control siRNA (C) for 72 h, and then incubated in the presence of 5 µM MG132 or DMSO. (B) Quantification of the densitometric... more
SCML2B stabilizes p21.(A) Western blot analysis of p21, p27, and actin, using whole cell extracts from U2OS cells treated with two different siRNAs (#1, #2) for SCML2 or a control siRNA (C) for 72 h, and then incubated in the presence of 5 µM MG132 or DMSO. (B) Quantification of the densitometric analysis for p21 and p27, from three different experiments performed as in (A). The intensity was corrected for the expression of actin, and the ratio of the levels after MG132 treatment relative to those of DMSO treatment is shown for each siRNA. *p<0.01 (C) U2OS cells were transfected with a control siRNA and plasmid (Control), with an siRNA against SCML2 and a control plasmid (si#2) or an siRNA against SCML2 and a plasmid expressing SCML2B resistant to the siRNA (si#2+SCML2B) for 48 h. Cells growing asynchronously (top panel) or released from an arrest in mitosis (5 h, middle panel, and 8 h, bottom panel) were then incubated with 25 µg/ml cycloheximide, and whole cells extracts were analyzed at different time points by Western blot with antibodies specific for p21. Densitometric quantification of the levels of p21 from two different experiments, each of them loaded in duplicate, is shown. The data were fitted to a one phase decay equation. less

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Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.

In PLoS Biol on 1 December 2013 by Lecona, E., Rojas, L. A., et al.

Fig.5.A
Fig.5.A showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
  • WB
Regulation of G1/S progression by SCML2.(A) Western blot analysis of nuclear extracts isolated from U2OS cells transfected with siRNA against either SCML2 (#2) or p21 (si p21) or a control siRNA (si C). Cells were collected every 2 h, as indicated, after release from a mitotic arrest induced with... more
Regulation of G1/S progression by SCML2.(A) Western blot analysis of nuclear extracts isolated from U2OS cells transfected with siRNA against either SCML2 (#2) or p21 (si p21) or a control siRNA (si C). Cells were collected every 2 h, as indicated, after release from a mitotic arrest induced with nocodazole. The extracts were analyzed using antibodies specific to USP7 (loading control), SCML2, CYCE2, CYCB2, CDK2, p27, and p21. (B) Densitometric analysis of the levels of p21 at different points after release from mitosis in cells treated with a control siRNA or specific for SCML2 (si #2). The expression was normalized to the levels of CDK2, and the mean of four different experiments is shown. (C) Western blot analysis of nuclear extracts isolated from U2OS cells transfected with a control siRNA and plasmid (Control), with an siRNA against SCML2 and a control plasmid (si #2) or an siRNA against SCML2 and a plasmid expressing SCML2B resistant to the siRNA (si #2+SCML2B). Cells were collected every 2 h, as indicated, after release from a mitotic arrest induced with nocodazole. The extracts were analyzed using antibodies specific to p21. Ponceau staining is shown as loading control. (D) Densitometric analysis of the levels of p21 at different time points after release from mitosis in two independent experiments. less

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Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.

In PLoS Biol on 1 December 2013 by Lecona, E., Rojas, L. A., et al.

Fig.4.E
Fig.4.E showing Western Blotting from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
  • WB
Role of SCML2 in cell cycle.(A) Western blot analysis of extracts prepared from U2OS cells treated with two different siRNAs (#1, #2) against SCML2 or a control siRNA (C), using antibodies specific to SCML2, CDK2, p27, and p21. (B) Cell-cycle distribution of U2OS cells after SCML2 knockdown using... more
Role of SCML2 in cell cycle.(A) Western blot analysis of extracts prepared from U2OS cells treated with two different siRNAs (#1, #2) against SCML2 or a control siRNA (C), using antibodies specific to SCML2, CDK2, p27, and p21. (B) Cell-cycle distribution of U2OS cells after SCML2 knockdown using the siRNAs indicated, measured by PI staining and FACS. *p<0.05, **p<0.01. (C–D) Cell-cycle distribution of U2OS cells after treatment with a control siRNA (Control), siRNA against either SCML2 (#2) or p21, or a combination thereof for SCML2 and p21 (C), or for SCML2 (#2) and a combination of SCML2 and CYCE2 (D), measured by PI staining and FACS. Statistical significance was determined with paired t tests. *p<0.05, **p<0.01, ***p<0.001. (E) Western blot analysis of extracts prepared from U2OS cells treated with siRNA against either SCML2 (#2), or p21 or CYCE2, or a combination thereof for SCML2 and p21, or for SCML2 and CYCE2, or a control siRNA (first column), using antibodies specific to SCML2, CYCE2, p21, CYCB2, p27, and USP7 (loading control). (F) U2OS cells were synchronized in mitosis with nocodazole after SCML2 knockdown, and the cell-cycle distribution after 16 h of release is compared. (G) Quantification of the proportion of cells in S phase at different time points after release from mitotic arrest from the experiment shown in Figure S8. *p<0.05. less

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Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.

In PLoS Biol on 1 December 2013 by Lecona, E., Rojas, L. A., et al.

Fig.4.A
Fig.4.A showing Western Blotting in a Homo sapiens (Human) sample from the publication: Polycomb protein SCML2 regulates the cell cycle by binding and modulating CDK/CYCLIN/p21 complexes.
  • WB
  • Homo sapiens (Human)
Role of SCML2 in cell cycle.(A) Western blot analysis of extracts prepared from U2OS cells treated with two different siRNAs (#1, #2) against SCML2 or a control siRNA (C), using antibodies specific to SCML2, CDK2, p27, and p21. (B) Cell-cycle distribution of U2OS cells after SCML2 knockdown using... more
Role of SCML2 in cell cycle.(A) Western blot analysis of extracts prepared from U2OS cells treated with two different siRNAs (#1, #2) against SCML2 or a control siRNA (C), using antibodies specific to SCML2, CDK2, p27, and p21. (B) Cell-cycle distribution of U2OS cells after SCML2 knockdown using the siRNAs indicated, measured by PI staining and FACS. *p<0.05, **p<0.01. (C–D) Cell-cycle distribution of U2OS cells after treatment with a control siRNA (Control), siRNA against either SCML2 (#2) or p21, or a combination thereof for SCML2 and p21 (C), or for SCML2 (#2) and a combination of SCML2 and CYCE2 (D), measured by PI staining and FACS. Statistical significance was determined with paired t tests. *p<0.05, **p<0.01, ***p<0.001. (E) Western blot analysis of extracts prepared from U2OS cells treated with siRNA against either SCML2 (#2), or p21 or CYCE2, or a combination thereof for SCML2 and p21, or for SCML2 and CYCE2, or a control siRNA (first column), using antibodies specific to SCML2, CYCE2, p21, CYCB2, p27, and USP7 (loading control). (F) U2OS cells were synchronized in mitosis with nocodazole after SCML2 knockdown, and the cell-cycle distribution after 16 h of release is compared. (G) Quantification of the proportion of cells in S phase at different time points after release from mitotic arrest from the experiment shown in Figure S8. *p<0.05. less

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Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.

In PLoS One on 31 July 2012 by Wang, H., Xu, Y., et al.

Fig.5.B
Fig.5.B showing Western Blotting in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
  • WB
  • Mus musculus (House mouse)
p27 downregulation in response to AKT activation is dependent on the nonconserved T157.(A) Sequences of human and murine p27. (B) Endogenous p27 expression in serum starved human CWR22Rv1 cells stimulated with IGF-1 for 0–6 hours. (C) Flag epitope tagged wild-type or T157A mutant human p27 expres... more
p27 downregulation in response to AKT activation is dependent on the nonconserved T157.(A) Sequences of human and murine p27. (B) Endogenous p27 expression in serum starved human CWR22Rv1 cells stimulated with IGF-1 for 0–6 hours. (C) Flag epitope tagged wild-type or T157A mutant human p27 expression vectors were transiently transfected into CWR22Rv1 PCa cells for 24 hours followed by overnight serum starvation. Cells were then treated with IGF-1 (10 ng/ml) as indicated, harvested after 4 or 8 hours, and lysates were immunoblotted for Flag-p27, pAKT, and β-tubulin (loading control). (D) TRAMP-C2 murine PCa cells were serum starved overnight and then IGF-1 stimulated as indicated. (E) TRAMP-C2 cells were transfected with Flag tagged human p27 expression vectors, serum starved overnight, and then treated with IGF-1 as in (C), followed by immunoblotting for Flag and pAKT. (F) Average Flag/actin ratios at 2 and 4 hours after IGF-1 stimulation (normalized to time 0) from three experiments were quantified. less

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Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.

In PLoS One on 31 July 2012 by Wang, H., Xu, Y., et al.

Fig.5.D
Fig.5.D showing Western Blotting in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
  • WB
  • Mus musculus (House mouse)
p27 downregulation in response to AKT activation is dependent on the nonconserved T157.(A) Sequences of human and murine p27. (B) Endogenous p27 expression in serum starved human CWR22Rv1 cells stimulated with IGF-1 for 0–6 hours. (C) Flag epitope tagged wild-type or T157A mutant human p27 expres... more
p27 downregulation in response to AKT activation is dependent on the nonconserved T157.(A) Sequences of human and murine p27. (B) Endogenous p27 expression in serum starved human CWR22Rv1 cells stimulated with IGF-1 for 0–6 hours. (C) Flag epitope tagged wild-type or T157A mutant human p27 expression vectors were transiently transfected into CWR22Rv1 PCa cells for 24 hours followed by overnight serum starvation. Cells were then treated with IGF-1 (10 ng/ml) as indicated, harvested after 4 or 8 hours, and lysates were immunoblotted for Flag-p27, pAKT, and β-tubulin (loading control). (D) TRAMP-C2 murine PCa cells were serum starved overnight and then IGF-1 stimulated as indicated. (E) TRAMP-C2 cells were transfected with Flag tagged human p27 expression vectors, serum starved overnight, and then treated with IGF-1 as in (C), followed by immunoblotting for Flag and pAKT. (F) Average Flag/actin ratios at 2 and 4 hours after IGF-1 stimulation (normalized to time 0) from three experiments were quantified. less

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Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.

In PLoS One on 31 July 2012 by Wang, H., Xu, Y., et al.

Fig.4.C
Fig.4.C showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
  • IHC
  • Mus musculus (House mouse)
p27 expression is rapidly induced by doxycycline prior to development of PIN.(A) p27 immunostaining in ventral prostate of double transgenic mice treated for 8 weeks with or without doxycycline, and in prostate from a nontrangenic normal control (higher power). (B) p27 and pAKT expression in doub... more
p27 expression is rapidly induced by doxycycline prior to development of PIN.(A) p27 immunostaining in ventral prostate of double transgenic mice treated for 8 weeks with or without doxycycline, and in prostate from a nontrangenic normal control (higher power). (B) p27 and pAKT expression in double transgenic ventral prostate after 2 weeks of doxycycline. (C) pAKT and p27 expression in ventral prostate of adult double transgenic mice treated with doxycycline for 2, 4, or 7 days. Sections shown are representative of ventral prostates from at least 3 mice at each time. less

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Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.

In PLoS One on 31 July 2012 by Wang, H., Xu, Y., et al.

Fig.4.B
Fig.4.B showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Doxycycline regulated induction of AKT in murine prostate drives proliferation independently of p27 cyclin dependent kinase inhibitor downregulation.
  • IHC
  • Mus musculus (House mouse)
p27 expression is rapidly induced by doxycycline prior to development of PIN.(A) p27 immunostaining in ventral prostate of double transgenic mice treated for 8 weeks with or without doxycycline, and in prostate from a nontrangenic normal control (higher power). (B) p27 and pAKT expression in doub... more
p27 expression is rapidly induced by doxycycline prior to development of PIN.(A) p27 immunostaining in ventral prostate of double transgenic mice treated for 8 weeks with or without doxycycline, and in prostate from a nontrangenic normal control (higher power). (B) p27 and pAKT expression in double transgenic ventral prostate after 2 weeks of doxycycline. (C) pAKT and p27 expression in ventral prostate of adult double transgenic mice treated with doxycycline for 2, 4, or 7 days. Sections shown are representative of ventral prostates from at least 3 mice at each time. less

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Pten dose dictates cancer progression in the prostate.

In PLoS Biol on 1 December 2003 by Trotman, L. C., Niki, M., et al.

Fig.5.C
Fig.5.C showing Immunohistochemistry in a Mus musculus (House mouse) sample from the publication: Pten dose dictates cancer progression in the prostate.
  • IHC
  • Mus musculus (House mouse)
Molecular Effects of Loss of Pten and Biological Comparison of All Generated Mouse Models(A) Ki-67 staining of AP lobe sections illustrates increasing proliferation with decreasing Pten levels (numbers are Ki-67-positive cells per 300 cells counted in percent; bars are 50 μm).(B) The phospho-Akt/... more
Molecular Effects of Loss of Pten and Biological Comparison of All Generated Mouse Models(A) Ki-67 staining of AP lobe sections illustrates increasing proliferation with decreasing Pten levels (numbers are Ki-67-positive cells per 300 cells counted in percent; bars are 50 μm).(B) The phospho-Akt/Akt ratio is sharply increased in the prostates of 10-wk-old Ptenpc2 animals, as shown by densitometric quantification of WB analysis.(C) AP staining with anti-phospho-Akt antibodies reveals strong plasma membrane localization of phospho-Akt and apparent reduction of p27 protein detection, whereas phospho-mTOR and phospho-threonine-FOXO3 antibodies show increased staining in Ptenpc2 versus wild-type mouse prostates. Bars are 50 μm.(D) Kaplan–Meier curve showing prostate enlargement as visualized by MRI. Progressive rates of mass increase for Ptenpc2 (median age, 4 mo), Ptenhy/− (median age, 7 mo), and Ptenpc1 (median age, 16 mo) mice are found. In contrast, no prostatic size irregularities were detected in Pten+/+ or Pten+/− mice.(E) Incidence of invasive CaP. Invasive CaP was defined as tumor cells disrupting the basal membrane of prostatic glands and growing into the surrounding stroma. Full penetrance was observed in both Ptenpc1 mice as well as in Ptenpc2 mice. In contrast, Ptenhy/− mice with a follow-up of more than 6 mo displayed only 25% incidence of invasive CaP. less

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