All lanes : Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody (ab23875)Lane 1 : Lysates prepared from HepG2 cells left unstimulated with 3% BSA-TBST buffer and no peptideLane 2 : Lysates prepared from HepG2 cells stimulated with Metformin with 3% BSA-TBST buffer and no peptideLane 3 : Lysates prepared from HepG2 cells stimulated with Metformin with 3% BSA-TBST buffer and the non-phosphopeptide corresponding to the immunogenLane 4 : Lysates prepared from HepG2 cells stimulated with Metformin with 3% BSA-TBST buffer and a generic phospho-threonine-containing peptideLane 5 : Lysates prepared from HepG2 cells stimulated with Metformin with 3% BSA-TBST buffer and the phosphopeptide immunogenLane 6 : Lysates prepared from HepG2 cells stimulated with Metformin and treated with lambda phosphatase with 3% BSA-TBST buffer Secondarygoat F(ab’)2 anti rabbit IgG HRP conjugate