Regulation of high glucose-induced NF-κB nuclear translocation and c-Src activity by Cx43 is independent of GJIC. GMCs were transfected with GFP-Cx43 or Flag-Cx43CT under the condition of normal glucose (NG; 5.5 mmol/L). After 48 h of transfection, GMCs were exposed to high glucose (HG; 30mmol/L) for 30 min. (A) Photomicrographs obtained after Lucifer yellow scrape-loading in GMCs transfected with GFP-Cx43 and Flag-Cx43CT (magnification 100×, upper panel). Phase contrast views are also provided (lower panel). Scale bar represents 100 μm. (B) Proteins were extracted for analysis of NF-κB p65 nuclear translocation by immunoblotting. Histone H1.4 and α-tubulin were measured by immunoblotting as a loading control. (C) c-Src was immunoprecipitated with an anti-c-Src antibody and Flag, phosphorylation of Tyr416 on c-Src (Y416-c-Src) and total c-Src were analyzed by immunoblotting. Experiments were performed at least three times with similar results. *P<0.05 vs. normal glucose-treated group, #P<0.05 vs. 30 mmol/L glucose-treated group.