Figure 2

p30 decreases the expression of proteins involved in S phase entry and progression. Hela cells were transduced for 48 h with GFP (control) or p30-myc expressing lentivirus. A) p30-myc expression was detected by western blot (WB) using anti-myc antibody or B) by indirect immunofluorescence (IF) staining. Nearly 100% of cells were transduced as shown with different magnifications. GFP-viral particles (HRCMV-GFP) were used as a control in the IF staining. C) WB analysis of PCNA protein was performed in the same experimental conditions. D) Quantification of E2F, cyclin E and p21waf mRNAs by real time PCR. E2F, Cyclin E and p21waf values were normalized to that of GAPDH which was used as an internal control. E) Extracts of Hela cells transduced with GFP or p30-myc virus were used for WB analyses of the endogenous E2F, cyclin E and p21 proteins. F) 293T cells were transfected with 6xE2F-Luciferase reporter plasmid either with HTLV-1 p30 or HTLV-2 p28 expression vectors. Histograms represent the values of three different experiments with their standard deviations. WB analyses showing equal amounts of p30 and p28 are present in the cell extracts.