Fig. 6: Metformin blocked the interaction of MET and Gab1 and thereby inhibited the phosphorylation of Gab1.

a H3122/Vec and H3122/HGF cells pretreated with alectinib (50 nmol/L) and/or metformin (5 mmol/L) for 48 h were lysed and then subjected to denaturing Co-IP with M2 beads followed by western blot analysis. Similar results were obtained in three independent experiments. b H2228/Vec and H2228/HGF cells pretreated with alectinib (500 nmol/L) and/or metformin (5 mmol/L) for 48 h were lysed and then subjected to denaturing Co-IP with M2 beads followed by western blot analysis. Similar results were obtained in three independent experiments. c H3122/HGF cells were treated with alectinib (50 nmol/L), JNJ-38877605 (10 nmol/L) and/or metformin (5 mmol/L) for 48 h. Cell lysates were harvested and subjected to denaturing Co-IP with M2 beads followed by western blot analysis. Similar results were obtained in three independent experiments. d H3122 cells were transfected with p-Gab1^Y627A-lentivirus, p-Gab1^Y627D-lentivirus, wild-type p-Gab1-lentivirus, and corresponding empty vector, and the phosphorylation levels of Gab1 were detected by western blot analysis. e H3122 cells were transfected with p-Gab1^Y627A-lentivirus, p-Gab1^Y627D-lentivirus, and wild-type p-Gab1-lentivirus and then treated with HGF (50 ng/mL) and alectinib (50 nmol/L) with or without metformin (5 mmol/L) for 48 h. Cell lysates were harvested and subjected to denaturing Co-IP with M2 beads followed by western blot analysis. Similar results were obtained in three independent experiments. Ale alectinib, Metf metformin, METi MET selective inhibitor JNJ-38877605, Vec negative control vector, NC empty vector, WT wild-type p-Gab1.