Fig. 8: ATAT1 mediates p27 regulation of autophagy in glucose-deprived cells.

A p27+/+ and p27−/− mCherry-eGFP LC3 MEFs were transfected with siRNA against ATAT1 or control siRNA. After 24 h, cells were starved for either glucose or aa for further 24 h. Scale bar 50 µm. B, C Graphs show the percentage of autophagosomes (yellow puncta) and autolysosomes (red puncta) in glucose-deprived cells (B) and amino acid-deprived cells (C), as described in (A). Number of analyzed images: Glucose starvation: p27+/+ siControl n = 31; p27+/+ siATAT1 n = 45; p27−/− siControl n = 38, p27−/− siATAT1 n = 49; Amino acid starvation: p27+/+ siControl n = 40; p27+/+ siATAT1 n = 34; p27−/− siControl n = 38, p27−/− siATAT1 n = 57 Statistical significance was determined by 2-way ANOVA followed by Bonferroni multiple comparison test. D Immunoblot showing the efficiency of siATAT1 by monitoring microtubule acetylation levels in p27+/+ and p27−/− MEFs. Membranes were probed for Ac-α-tubulin, p27 and β-tubulin as loading control.