Fig. 5

HG/PA down-regulates WWP2 through JNK activation. A Representative Western blots and quantification of p-JNK protein expression levels in HUVECs with or without HG/PA treatment. Values are shown as mean ± SD (**P < 0.01, ***P < 0.001, two-tailed unpaired Student t-tests). B Relative mRNA levels of WWP2 in HUVECs treated with JNK inhibitor (SP600125) or JNK activator (Anisomycin), with or without HG/PA. C, D Representative Western blots (C) and quantification (D) of WWP2 and p-JNK protein expression levels in HUVECs treated with JNK inhibitor (SP600125) or JNK activator (Anisomycin), with or without HG/PA. E The proliferation and viability of HUVECs was tested using the CCK-8 colorimetric assay. HUVECs were transfected with WWP2 siRNA or a negative control, and treated with JNK inhibitor (SP600125), with or without HG/PA. F, G Representative Flow cytometry analysis (F) and quantitative analysis (G) of apoptosis in HUVECs. H, I Representative Western blots (H) and quantification (I) of WWP2, p-JNK, Cleaved PARP1 and Cleaved caspase-3 protein expression levels in HUVECs. HUVECs were transfected with WWP2 siRNA or a negative control, and treated with JNK inhibitor (SP600125), with or without HG/PA. Values are shown as mean ± SD (*P < 0.05, **P < 0.01, ***P < 0.001, NS = not significant, one-way ANOVA with Tukey’s multiple comparison post-hoc test). CCK-8, Cell Counting Kit-8; Ctrl, control; HG/PA, high glucose/palmitic acid; HUVECs, HUVECs, Human umbilical vein endothelial cells; NC, negative control; p-JNK, phosphorylation of JNK; siRNA, small interfering RNA