Extended Data Fig. 2: AMPK plays a pivotal role in mTORC1 glucose sensing.
a, IB analysis of WCLs derived from WT and AMPKα1/2 double knockout (DKO) HEK 293 cells. The cells were deprived of glucose for 60 min and restimulated with glucose for 10 min as indicated. b, IB analysis of WCLs derived from WT and AMPKα1/2 DKO HEK 293 T cells. The cells were deprived of glucose or amino acids for 60 min and restimulated with glucose or amino acids for 10 min as indicated. c, IB analysis of WCLs derived from WT and Ampkα1/2 DKO MEFs. The cells were treated as in (a). d, IB analysis of WCLs derived from WT, AMPKα1/2 DKO, AMPKα1 and AMPKα2 reconstituted AMPKα1/2 DKO U2OS cells. The cells were treated as in (a). e,f, HEK 293 cells were treated with or without A-769662 (100 μM) for 60 min, and the co-localization of mTORC1 (Red) and LAMP1 (green) was analyzed via immunofluorescence (e). Scale bar, 10 μm. The imaging data were quantified (f). n = 17, 20. P = 0.00058. Data are the mean ± s.d., two-tailed t-test. NS, not significant, ***P < 0.001. Representative image shown, a, b and d, n = 2; c, n = 3.
