Fig. 3: WWP2 overexpression alleviates apoptosis under the inducement of Dox in Jurkat cells while knockout WWP2 enhances this effect.

A–C Concentration gradient of Dox was set for western blot and cell viability assay (0, 0.05, 0.1,0.2, 0.4 μM) and for flow cytometry analysis (0, 0.025, 0.05, 0.075, 0.1 μM). A Flow cytometry annexin-FITC/PI was used to assess apoptosis rate of Jurkat cells in different concentration of Dox. Data were shown as mean ± SD (*p < 0.05, **p < 0.01; one-way ANOVA with Bonferroni’s post-hoc test). B Western blot was used to evaluate apoptosis protein (Bax and cleaved-C3) expression of Jurkat cells in different concentration of Dox. Data were shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Bonferroni’s post-hoc test). C CCK-8 assay was used to assess the viability of Jurkat cells in different concentration of Dox. Data were shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA with Bonferroni’s post-hoc test). D–F Jurkat cells were transfected with HA-WWP2 or HA-control plasmids under the inducement of Dox for western blot and cell viability assay (0.2 μM) and for flow cytometry (0.075 μM). D Flow cytometry annexin-FITC/PI was used to assess apoptosis rate of Jurkat cells. Data were shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001; unpaired Student’s t test, two-way ANOVA with Bonferroni’s post-hoc test). E Western blot was used to evaluate apoptosis protein (Bax and cleaved-C3) expression of Jurkat cells. Data were shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001; unpaired Student’s t test, two-way ANOVA with Bonferroni’s post-hoc test). F CCK-8 assay was used to assess the viability of Jurkat cells. Data were shown as mean ± SD (*p < 0.05, **p < 0.01, ***p < 0.001; two-way ANOVA with Bonferroni’s post-hoc test). G–I Jurkat cells were infected with WWP2 knockout or control lentivirus under the inducement of Dox for western blot and cell viability assay (0.2 μM) and for flow cytometry (0.075 μM). G Flow cytometry, H western blot and I CCK-8 assays were performed as described above.