Fig. 3: Atg7 senses ATP levels and promotes PDCD4 phosphorylation at Ser67.
a Western blot detection of PDCD4 and p-PDCD4 Ser67 in HEK293 Atg7-WT, Atg7-KO, and HEK293 Atg7-KO cells transfected with Flag-Atg7 with or without 2DG (5 mM) and oligomycin (2.5 μM) stimulation for four hours. b Interaction between endogenous AKT and PDCD4 in HEK293 Atg7-KO and Atg7-WT cells. c HEK293 cells were transfected with GFP-Atg7, GFP-Atg7D476AQ398A, or control vector and the indicated plasmids. Co-immunoprecipitation assay was performed to detect the interaction between Flag and AKT. d HEK293 Atg7-KO cells were transfected with Flag-Atg7, Flag-Atg7D476AQ398A, or control vector. After pretreated with CQ (10 μM) in complete medium for 12 h and then treated or untreated with H2O2 (500 μM) or 2DG (5 mM) and oligomycin (2.5 μM) stimulation for four hours then collected for Western blot analysis. e HEK293 Atg7-KO cells were transfected with GFP-Atg7, GFP-Atg7D476AQ398A, or control vector and the indicated plasmids. Co-immunoprecipitation assay was performed to detect the interaction between Flag and AKT after 2DG (5 mM) and oligomycin (2.5 μM) treatment for four hours. f HEK293 cells were transfected with GFP-Atg7, GFP-Atg7D476AQ398A, or control vector and the indicated plasmids. Co-immunoprecipitation assay was performed to detect the interaction between GFP and AKT or Flag.
