Fig 7.
Metformin mediated its action via LKB1. (A) LKB null (LKB2/2) and wild-type (LKB+/+) mefs were treated with metformin (5–10 mM) and analysed by immunoblot for AMPK and ACC phosphorylation and b actin for equal protein loading. (B) LKB2/2 and wild-type mefs were treated with metformin with indicated concentrations for 24 hrs. After overnight fixation and propidium iodide staining cells were flow-sorted. The data are representations of two separate experiments. (C) Immunoblot analysis revealed down-regulation of LKB1 in untransfected (C), siRNA (Si) and non-target siRNA control (NT) transfected A2780 cells at 48 hrs. (D) After 48 hrs transfection, similar sets were treated with metformin for 12 hrs and processed for immunoblot for p-ACC and total ACC. (E) A2780 untransfected (C), siRNA (Si) and non-target siRNA control (NT) transfected cells were treated with indicated concentrations of metformin for 48 hrs and assessed for proliferation by MTT. **P < 0.01, siRNA (Si) compared to C and NT.