Fig. 8

DDX3X is essential for the regulation of WWP2 on endothelial injury. A, B Representative Western blots (A) and quantification (B) of DDX3X protein expression levels in HUVECs. HUVECs were transfected with three siRNA sequences of DDX3X to evaluate the efficiency of DDX3X silencing. Values are shown as mean ± SD (***P < 0.001, one-way ANOVA with Dunnett’s multiple comparison post-hoc test). c CCK-8 colorimetric assay was used to test the proliferation and viability of HUVECs in each indicated group. HUVECs were transfected with DDX3X siRNA or a negative control, and transfected with WWP2 siRNA or a negative control, with or without HG/PA treatment. D, E Representative Flow cytometry analysis (D) and quantitative analysis (E) of HUVECs apoptosis in each indicated group. F, G Representative Western blots (F) and quantification (G) of WWP2, DDX3X, Cleaved PARP1 and Cleaved caspase-3 protein expression levels in HUVECs of each indicated group. Values are shown as mean ± SD (*P < 0.05, ***P < 0.001, ###P < 0.001, NS = not significant, two-way ANOVA with Bonferroni’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; siRNA, small interfering RNA