Figure 3

CsH increases the efficiency of multi-vector transduction of murine HSCs in vitro. HSCs were sorted from murine BM, transduced with a mix of LVs designed to express ZsGreen, mCherry or RFP670 fluorescent reporters, incubated with LVs and CsH/DMSO for 24 hours, and cultured for 4 days. The percentage of cells expressing fluorescent reporters was determined using flow cytometry (A) Representative FACS plots for reporter-derived fluorescence in uninfected (no LVs, left) and triple-infected (LV mix) cells. ZsGreen and RFP670 were gated first (left plots), and then the expression of mCherry in the ZsGreen⁺RFP⁺ population was assessed (right plots). The results from DMSO-treated controls and samples treated with CsH are shown in the top and bottom plots, respectively. Separate plots showing the total fraction of mCherry-positive cells are presented in Supplementary Fig. 3. Data were collected from four independent experiments (n = 4), and triplicate technical replicates were used within the experiments. (B–E) Quantification of each single colour (B–D) and of the triple-transduced cells (E). MOI values: ZsGreen – 50, mCherry – 40, and RFP670–25. The results are presented as the means ± SDs. Statistical significance was calculated using an unpaired t-test. *p ≤ 0.05, **p ≤ 0.01.