Fig. 8. p62 acetylation promotes the clearance of poly-Ub proteins.

a Sequestration of p62 bodies by autophagosomes. p62-KO HEK293 cells stably expressing Flag-tagged WT p62, p62-2KQ or p62-2KR were transfected with GFP-LC3B and subjected to starvation. The cells were then stained with anti-Ub and imaged by confocal microscopy. Scale bars, 10 µm. b The percentage of GFP-LC3B puncta in a that were also positive for ubiquitin. The data are presented as mean ± S.E.M., n = 30 cells. Two-tailed t-test, ***p < 0.001. c, d The level of poly-Ub proteins in p62-KO HEK293 cells stably expressing Flag-tagged WT p62 or the p62 mutants. The cells were starved with or without Baf-A1 for 24 h and analyzed by western blot using anti-Ub (c). The ratios of the intensity of ubiquitylated proteins in Baf-A1-treated samples over that in Baf-A1-untreated samples are presented in d. The data are presented as mean ± S.E.M. of three independent experiments. Two-tailed t-test, *p < 0.05, ***p < 0.001. e, f The degradation of Flag-p62 in p62-KO HEK293 cells stably expressing Flag-tagged WT p62 or the p62 mutants upon cell starvation. Flag-p62 was detected by western blot using anti-Flag (e), and the quantifications are presented as mean ± S.E.M. of three independent experiments (f). Two-tailed t-test, ***p < 0.001. g Viability of p62-KO HEK293 cells stably expressing Flag-tagged WT p62 or the p62 mutants after cell starvation with or without Baf-A1 for 48 h. The data are presented as mean ± S.E.M. of five independent experiments. Two-tailed t-test, ***p < 0.001. Source data are provided as a Source Data file.