Fig. 6: HDAC inhibition restores autophagosomes positioning in p27−/− cells. | Cell Death & Disease

Fig. 6: HDAC inhibition restores autophagosomes positioning in p27−/− cells.

From: p27 controls autophagic vesicle trafficking in glucose-deprived cells via the regulation of ATAT1-mediated microtubule acetylation

Fig. 6

A LC3B, LAMP2 and Ac-α-tubulin immunostaining of p27+/+ and p27−/− MEFs glucose deprived for 48 h ± 0.2 µM TSA for 1 h. Scale bars are 50 µM. B Acetylated tubulin fluorescence intensity was measured in cells glucose-starved for 48 h. Values were normalized to 48 h glucose-starved p27+/+ cells. At least 160 cells were analyzed per condition in each experiment from n = 3 independent experiments. C Immunoblotting for Ac-α-tubulin and tubulin in cells as described in A. Actin was used as loading control. * denotes remaining Ac-α-tubulin signal after membrane stripping. D Percentage of p27+/+ and p27−/− MEFs glucose deprived for 48 h ± 0.2 µM TSA for 1 h with >50 % of LC3B vesicles in the perinuclear region. Autophagosome distribution was evaluated based on a percentage of LC3 integrated density in perinuclear area (designed by region of interest [ROI]) versus LC3 integrated density in the whole cell. At least 91 cells were analyzed per condition. n = 3 independent experiments. B, D Bar graphs show mean ± SEM. Statistical significance was determined using 2-way ANOVA followed by Bonferroni multiple comparison test.

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