Extended Data Fig. 5: Phosphorylation of S155 on WDR24 inhibits the activation of mTORC1 induced by glucose.
a, WDR24 KO HeLa cells re-introduced with indicated constructs were deprived of glucose for 60 min and restimulated with glucose for 10 min as indicated. b, Cell size histogram of WDR24 KO HeLa cells reconstituted with WT and WDR24 S155D by FACS. c, WDR24 KO HeLa cells reconstituted with indicated constructs were plated for 8 d for the colony-formation assays. Data are shown as the mean ± s.d. of n = 3 independent experiments (bottom). P = 0.0004. two-tailed t-test. ***P < 0.001. d, IB analysis of WCLs of HEK 293 cells starved of glucose or/and amino acids for 1 h, and restimulated with amino acids for 10 min. e, WDR24 KO HEK 293 cells re-introduced with indicated constructs were deprived of amino acids for 60 min and restimulated with amino acids for 10 min as indicated before IB analysis. f, WDR24 KO HeLa cells were re-introduced with indicated constructs. The cells were treated as in (e). g,h, WT and AMPKα1/2 DKO HEK 293 (g) or 293 T (h) cells were starved of amino acids or amino acids and glucose together for 60 min, then stimulated with amino acids for 10 min. i, Working model to show how AMPK regulates mTORC1 amino acid sensing under energy stress conditions. Representative image shown, a and d, n = 3; e, f, g and h, n = 2.
