Fig. 6.

Metformin can accelerate the phosphorylation of AMPK in HeLa cells (A) Phosphorylation of AMPK was confirmed in metformin-treated HeLa cells. (A) HeLa (without LKB1 gene) and (B) HEC-1-A (with LKB1) cells were treated with metformin, respectively. HEC-1-A cell lines compared to HeLa cervical cells. Then, whole cells were collected and added with lysis buffer. After harvest by centrifugation, equal amounts of protein (20 μg) were separated on 8-12% SDS-PAGE, followed by western blot analysis with indicated specific antibodies (AMPK/phosphorylated AMPK, mTOR/phosphorylated mTOR, p70S6K/ phosphorylated p70S6K and 4E-BP1/phosphorylated 4E-BP1). Non-phosphorylated form (indicated AMPK, mTOR, p70S6K and 4E-BP1) included to verify equal loading of the samples, respectively. All data were conducted at least three times with consistent and similar results.