Research Article

SUMO E3 Ligase PIASy Mediates High Glucose-Induced Activation of NF-κB Inflammatory Signaling in Rat Mesangial Cells

Figure 2

The phosphorylation and sumoylation of IKKγ were induced by high glucose. (a) The protein expression of p-IKKγ and IKKγ in lysates of GMCs was detected after high glucose challenge for various times and various glucose concentrations by Western blotting. (b) IKKγ sumoylation was detected by immunoprecipitation (IP) with anti-IKKγ antibody followed by Western blotting with anti-SUMO1 or anti-SUMO2/3 antibody. IKKγ was conjugated with SUMO in GMCs. IgG (H) marks the IgG heavy chain. (c) GMCs were treated with 30 mmol/L high glucose or mannitol for 24 h. Anti-IKKγ immunoprecipitates were subjected to immunoblotting with anti-SUMO1 or anti-SUMO2/3 antibody to detect IKKγ and SUMO-IKKγ proteins; normal IgG antibody was used as a negative control. IKKγ that was sumoylated by SUMO1 or SUMO2/3 was induced by high glucose. The gray graphs confirmed these trends. Data are expressed as mean ± SD (). compared with the NC group, # compared with the 30 mmol/L high glucose stimulation group.