Fig. 4: TMEM189 knockdown increases ULK1 stability and kinase activity.

a, c Western blot analysis of endogenous indicated protein in cell lysates extracted from the stable shcontrol- or shTMEM189-expressing HEK293T cells. b, d Quantification of amounts of indicated protein relative to GAPDH in cells. Average value in shcontrol-infected cells was normalized as 1. Data are means ± SD of results from 3 experiments. e HEK293T cells were cotransfected with indicated plasmids for 24 h, then incubated with cycloheximide (CHX, 100 μg/mL) for indicated time. The levels of GFP-ULK1 and TMEM189 were detected by western blotting. f Quantification of GFP-ULK1 proteins (ratio to ACTB) in cells treated as in (e). g HEK293T cells were transfected with indicated plasmids for 24 h, incubated in cycloheximide (CHX, 100 μg/mL) for indicated time. The levels of endogenous ULK1 were detected by western blotting. h The stable shcontrol- or shTMEM189-expressing HeLa cells were incubated with CHX (100 μg/mL) for indicated time. The levels of endogenous ULK1 were detected by western blotting. i Quantification of endogenous ULK1 proteins (ratio to ACTB) in cells treated as in (h). J The stable shcontrol- or shTMEM189-expressing HEK293T cells were incubated with EBSS for indicated time. The levels of endogenous ULK1, TMEM189 and GAPDH were detected by western blotting.