SUMO E3 Ligase PIASy Mediates High Glucose-Induced Activation of NF-κB Inflammatory Signaling in Rat Mesangial Cells
The expression of PIASy and SUMO isoforms (SUMO1, SUMO2/3) was induced by various times and various glucose concentrations. GMCs were treated with 30 mmol/L high glucose for 6, 12, 24, 48, and 72 h or the indicated concentrations of glucose or mannitol for 24 h; the protein expression of PIASy (a) and SUMO isoforms (SUMO1, SUMO2/3) (b) in lysates of cells was detected by Western blotting. Data were normalized with respect to β-actin and are expressed as mean ± SD (). The gray graphs confirmed these trends. Immunofluorescence was performed to determine intensity and subcellular localization of PIASy and SUMO1 (c) or SUMO2/3 (d) after GMC treatment with 30 mmol/L high glucose or mannitol for 24 h (400x). The merged images of PIASy and SUMO1 or SUMO2/3 stainings in each group were shown, and DAPI was used to stain the nucleus in the cells. The values of semiquantitative analysis for average intensity were assessed, and the gray graphs confirmed these trends. compared with the NC group, # compared with the 30 mmol/L high glucose stimulation group.
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