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Axonal autophagic vesicle transport in the rat optic nerve in vivo under normal conditions and during acute axonal degeneration.

In Acta Neuropathol Commun on 29 May 2024 by Luo, X., Zhang, J., et al.

Fig.4.A

Changes of autophagic pathways and motor proteins in the rat optic nerve 6 h after crush lesion. A Representative immunoblots of LC3, p62, Kif5, Dynein, p150Glued, Cathepsin D, and GAPDH 6 h after crush injury in the areas of the optic nerve 1 mm proximal and distal to the lesion site compared wi... more

Changes of autophagic pathways and motor proteins in the rat optic nerve 6 h after crush lesion. A Representative immunoblots of LC3, p62, Kif5, Dynein, p150Glued, Cathepsin D, and GAPDH 6 h after crush injury in the areas of the optic nerve 1 mm proximal and distal to the lesion site compared with the contralateral side. B–H Quantifications of the band intensities of LC3 II:LC3 I, p62, Kif5, Dynein, p150Glued, and Cathepsin D normalized to GAPDH as loading control. Error bars represent Mean ± SEM. 3 independent experiments were included. *P < 0.05; **P < 0.01 N.S. no significant difference, according to one-way ANOVA and Tukey multiple comparisons test less

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YTHDF2 facilitates aggresome formation via UPF1 in an m6A-independent manner.

In Nat Commun on 6 October 2023 by Hwang, H. J., Park, T. L., et al.

Fig.8.A

YTHDF2 increases an association between dynein motor protein and the CED-UPF1-misfolded polypeptide complex.a IPs of polypeptidyl-puro. HEK293T cells were either undepleted or depleted of endogenous YTHDF2. Two days later, the cells were treated with MG132 for 12 h and puromycin for 1 h before ha... more

YTHDF2 increases an association between dynein motor protein and the CED-UPF1-misfolded polypeptide complex.a IPs of polypeptidyl-puro. HEK293T cells were either undepleted or depleted of endogenous YTHDF2. Two days later, the cells were treated with MG132 for 12 h and puromycin for 1 h before harvesting. Then, the cell extracts were subjected to IPs using α-puromycin antibody. The cell lysate before IP was serially diluted three-fold and loaded in the three leftmost lanes to demonstrate that western blotting is semiquantitative in our experimental conditions. Relative intensities of bands in western blots of proteins obtained after IP in undepleted cells were arbitrarily set to 1.0; n = 3. b IPs of FLAG-CTIF. As performed as shown in a, except that the cells were not treated with puromycin and were subjected to IPs using α-FLAG antibody. Relative band intensities of proteins after IP in undepleted cells were arbitrarily set to 1.0; n = 3. c Proposed model for YTHDF2-facilitated aggresome formation. Misfolded polypeptides tend to form small cytoplasmic aggregates, which are recognized by eEF1A1, a component of CED complex. Then, the CED complex with misfolded polypeptides recruits UPF1, which increases the frequency and fidelity of movement of the complex toward aggresome via a dynein-mediated and microtubule-dependent retrograde manner. Concomitantly, YTHDF2 is recruited to the complex via its interaction with UPF1, increases the binding of the complex to dynein motor protein and, consequently, promotes the dynein-mediated retrograde transport. The molecular mechanism by which YTHDF2 promotes the association between dynein motor protein and the CED complex with misfolded polypeptides remains unknown. See the Discussion section for more details. less

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

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