High glucose induces dissociation of Cx43 and c-Src, and promotes interaction between c-Src and IκB-α. GMCs were incubated in normal glucose (NG; 5.5 mmol/L) or high glucose (HG; 30 mmol/L) for the indicated times. (A) Cx43 was immunoprecipitated with an anti-Cx43 antibody and c-Src was analyzed by immunoblotting. white triangles=Cx43; black squares=c-Src. (B) Phosphorylation of tyr416 on c-Src (Y416-c-Src) and total c-Src were analyzed by immunoblotting. α-Tubulin was measured by immunoblotting as a loading control. (C) IκB-α was immunoprecipitated with an anti-IκB-α antibody and Tyr-phosphorylation of IκB-α and c-Src were analyzed by immunoblotting. black circle= p-Tyr; black squares=c-Src; black triangles=IκB-α. (D) Ser-phosphorylation of IκB-α and total IκB-α were analyzed by immunoblotting. black circle=p-Ser-IκB-α; black triangles=IκB-α. α-Tubulin was measured by immunoblotting as a loading control. Experiments were performed at least three times with similar results. *P<0.05 vs. normal glucose-treated group.