Fig. 5

ATIC knockdown inhibits mTOR-S6 K1-S6 signaling through activating AMPK. a, Expression of ATIC was detected as in Fig. 1A. The AICAR level in 12 HCC cancer tissues was determined by ELISA assay. The expression of ATIC and AICAR level in patient #1 was set as 1 respectively. Correlation of relative expression of ATIC and AICAR level in 12 HCC patients was analyzed. b, cells were infected with indicated shRNAs for 72 h, followed by ELISA assays analysis AICAR level. The relative AICAR level was plotted. The AICAR level in shScr group was set as 1. ***, p < 0.001. c-d, HepG2 or Huh-7 cells were treated with indicated shRNAs (c) or AICAR (d) for 72 h. Status of AMPK was analyzed. e, cells were transfected with indicated shRNAs for 72 h. Status of mTOR-S6 K1-S6 signaling was analyzed. f-g, cells were transfected with indicated shRNA, 24 h later, cells were treated with 10 μM C-C (F) or AMPK shRNA (G) for 48 h. Status of AMPK-mTOR-S6 K1-S6 signaling was analyzed. h-i, HepG2 cells were infected with indicated shRNAs then treated with 10 μM C-C (h) or AMPK shRNA (i). mTOR were immunoprecipitated from these cells respectively. The immunoprecipitates were incubated with S6 K1 in a kinase assay system, followed by western blot analysis. (i, right panel), the efficiency of AMPK knockdown was determined by western blot