Fig. 7.

Effect of FACT knock down on early steps of HIV-1 derived lentiviral vectors in HEK293T cells. SSRP1 encoding gene knock down was conducted essentially as previously described [31] by performing two successive siRNA transfections (see “Methods” section). The amount of SSRP1 protein monitored by western blotting is reported in (a). This method allowed us to achieve more than 80% of apparent SSRP1 extinction. After knock down, FAIRE analyses were performed as indicated in “Methods” section and the results obtained in cells depleted for SSRP1 are reported in (b). The early steps of replication of lentiviral vectors carrying the GFP encoding gene were determined by measuring GFP-positive cells with flux cytometry (c) and quantification of the viral DNA populations at 0–48 h post-transduction using quantitative PCR (data obtained after a 20 nM siRNA treatment are reported in d for the 24 and 48 h time points, see full time course analysis in Additional file 7: Figure S7). Data obtained with the interferin transfector agent alone and control siRNA are also reported in the figure. All values are shown as the mean ± standard deviation (error bars) of four independent sets of experiments. The p-values were calculated by Student’s t-test and are shown as *p < 0.05 and **p < 0.005 to represent the probability of obtaining significant differences compared with untreated conditions