Fig. 4

ARF4 is an anti-apoptotic substrate for USP7

A Interaction between USP7 and ARF4 in SHG-140 was determined using the Co-IP assay. B SHG-140 cells were transfected with different shUSP7 for 24 h and ARF4 expression was determined by western blotting, n = 3. C. Expression of ARF4 was determined by western blotting after treatment of SHG-140 with different concentrations of P5091 for 48 h, n = 3. D, E Changes in apoptosis after overexpression of ARF4 in SHG-140 and T98G cells treated with P5091 (2 µM) for 48 h or transfected with shUSP7 for 24 h were detected by flow cytometry, n = 3. Q3 (Annexin V-FITC + PI-) subpopulation was considered early-stage apoptosis and Q2 (Annexin V-FITC + PI+) was late-stage apoptosis or necrosis. Cell proportion comparisons between groups were done in both early- and late-stage subpopulation separately and in total. F-I Changes in apoptotic proteins after overexpression of ARF4 in SHG-140 and T98G cells treated with P5091 (2 µM) for 48 h or transfected with shUSP7 for 24 h were observed by western blotting analysis, n = 3. Statistics are expressed as mean ± S.E.M., ^#P = NS, *P < 0.05,**P < 0.01,***P < 0.001, or ****P < 0.0001