Figure S1 - Insulin Promotes Glucose Consumption via Regulation of miR-99a/mTOR/PKM2 Pathway

Insulin induced phosphorylated mTOR (p-mTOR) expression levels, while overexpression of miR-99a and rapamycin treatment inhibited insulin-induced p-mTOR levels. (A) HepG2 and HL7702 cells were starved in serum-free medium for 18 h, then treated with insulin (200 nM) for 6 h. Cells were collected and subjected to immunoblotting using antibodies against p-mTOR and GAPDH. (B) The cells were transfected with miR-99a (+) or miR-SCR (–) precursor, and cultured for 60 hours, then treated with or without insulin as above. The expression levels of p-mTOR and GAPDH were analyzed by immunoblotting. (C) Cells were pretreated with rapamycin (5 nM) for 30 min, followed by insulin treatment for 6 h. The protein levels of p-mTOR and GAPDH were detected by immunoblotting. (PPTX)