c-Src regulates high glucose or Cx43-induced NF-κB p65 nuclear translocation. (A) GMCs were preincubated and maintained in 10 μM PP2 (c-Src inhibitor) or 10 μM PP3 (inactive analogue) for 30 min and until the end of the experiment. Cells were then incubated in normal glucose (NG; 5.5 mmol/L) or high glucose (HG; 30 mmol/L) for 30 min. Proteins were extracted for analysis of NF-κB p65 nuclear translocation by immunoblotting. (B) GMCs were transfected with Cx43-siRNA in normal glucose (NG; 5.5 mmol/L). After 48 h of transfection, GMCs were co-incubated with PP2 or PP3 (10 μM) for 30 min. Proteins were then extracted for analysis of Cx43 expression and NF-κB p65 nuclear translocation by immunoblotting. (C) GMCs were co-incubated with 10 μM PP2 (c-Src inhibitor) or 10 μM PP3, maintained in high glucose for 24 h and then the proteins were extracted for analysis of FN, ICAM-1, and TGF-β1 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. **P<0.05 vs. Cx43-siRNA transfected group.