FIGURE 4.

Podocyte VEGF^KD downregulates nephrin in diabetic and eNOS^−/−:VEGF^KD mice: (A) WB: show nephrin downregulation in eNOS^−/−:VEGF^KD + dox and diabetic kidneys (Brown-Forsythe ANOVA, p = 0.047), no significant difference was detected between DM-VEGF^KD − dox and + dox (Welch’s t-test) ; (B) IHC: nephin IF signals are clearly decreased in glomeruli from eNOS^−/−:VEGF^KD (+dox) and DM-VEGF^KD (+dox) kidneys; (C) WB: podocin decreased in eNOS^−/−:VEGF^KD (+dox) and diabetic kidneys (Brown-Forsythe ANOVA, p = 0.0001), no significant difference was detected between DM-VEGF^KD − dox and + dox (Welch’s t-test); (D) WB: VEGFR2 decreased in eNOS ^−/−:VEGF^KD (+dox) and diabetic kidneys (Brown-Forsythe ANOVA, p = 0.015) but differences (+dox vs. - dox) were not significant; (E) WB: significant β3-integrin upregulation was detected in eNOS^−/−:VEGF^KD (+dox) kidneys (Brown-Forsythe ANOVA, p = 0.03). Scale bars = 50 μm, + dox = VEGF^KD induction with doxycycline.