Fig. 6
From: Polyubiquitin chain-induced p62 phase separation drives autophagic cargo segregation
Phosphorylation of S403 promotes polyubiquitin chain-induced phase separation, p62 body formation and autophagic degradation. a The indicated GFP-p62 constructs were expressed in Atg12−/− cells and observed by confocal microscopy. Scale bar, 5 µm. b Cells from a were quantified for the number of p62 bodies. In all, >45 cells from a were assessed blind and quantified. Error bars indicate SD (n = 3). c Cells from a were quantified for the area of p62 bodies. In total, >38 cells from a were assessed blind and quantified. Error bars indicate SD (n = 3). d NRK cells transiently expressing the indicated GFP-p62 constructs were starved for the indicated time. Cell lysates were analyzed by western blot with the indicated antibodies. e Quantification of the indicated western blots in d. f The indicated mutated recombinant mCherry-p62 proteins were mixed with linear Ubx8 and the reaction was visualized by confocal microscopy. mCherry-p62, 5 µM; linear Ubx8, 2.5 µM. Scale bar, 5 µm
