Figure 5: POH1 increases Survivin and FOXM1 expression by regulating E2F1.
From: POH1 deubiquitylates and stabilizes E2F1 to promote tumour formation
(a) Real-time RT–PCR analysis of the mRNA levels of Survivin and FOXM1 in cells transfected with control siRNA or POH1 siRNA. Data are mean±s.d. (by t-test analysis, P values are shown in the graph, n=3) (b) Human liver tumour cells untransfected (parental) or transfected with control siRNA or POH1 siRNA-1 and -2 were harvested at 72 h. The cell lysates were analysed using the indicated antibodies. (c) PLC/PRF/5 and SMMC-7721 cells transduced with lentivirus containing POH1-Flag and the control were analysed by immunoblotting using the indicated antibodies. (d) Cells overexpressing POH1-Flag or control cells were transfected with control siRNA or siRNA against E2F1 for 48 h. Cell lysates were analysed by immunoblotting using the indicated antibodies. (e,f) PLC/PRF/5 cells overexpressing E2F1 were transfected with control siRNA or POH1 siRNA, 72 h after transfection, cell lysates were analysed by immunoblotting (e); cells were cultured for 10 days then stained with crystal violet. Representative data from the colony-formation assays are presented, n=3 (f). (g,h) PLC/PRF/5 cells overexpressing Survivin were transfected with control siRNA or POH1 siRNA, and cell lysates were analysed by immunoblotting (g); representative data from the colony-formation assays are presented, n=3 (h).
