Figure 3

Cx43 regulates high glucose-induced NF-κB nuclear translocation. (A) GMCs were transfected with Cx43-siRNA or GFP-Cx43 in normal glucose (NG; 5.5 mmol/L). After 48 h of transfection, GMCs were exposed to high glucose (HG; 30 mmol/L) for 30 min. Proteins were then extracted for analysis of Cx43 expression and NF-κB p65 nuclear translocation by immunoblotting. (B) Immunofluorescence images of GFP were captured under a laser scanning confocal microscope to assess the transfection efficiency of GFP-Cx43. Green fluorescence indicates GFP. Scale bar represents 100 μm (magnification 100×, a). Immunofluorescence images stained doubly with anti-Cx43 antibody and anti-NF-κB p65 antibody were captured under a laser scanning confocal microscope (magnification 400×, b). High glucose and Cx43-siRNA-transfection enhance NF-κB p65 nuclear translocation and GFP-Cx43-transfection inhibits high glucose-induced NF-κB p65 nuclear translocation. Red fluorescence indicates localization of NF-κB p65. Green fluorescence indicates localization of Cx43. Blue fluorescence indicates nuclei. Scale bar represents 20 μm. 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.