Fig. 5

EG-expressing FL and CB cells have a growth dependency on IL3 and SCF human cytokines. A Experimental design: CB CD34⁺ cells were transduced with CTL/EG-GFP lentiviral vectors and cultured for 21 days in the indicated cytokine conditions. B Percent of GFP⁺ cells during the culture for CTL (upper panel) and EG (lower panel) cells. Mean ± SEM of 3 biological replicates is shown. C Example of flow cytometry dot plots showing the percent of CD33⁺KIT⁺ cells obtained after 21 days of culture in the different cytokine conditions. D Cumulative cell numbers of CD33⁺KIT⁺ cells in GFP⁺ CB EG cells at day 21 of culture. Mean ± SEM of 3 biological replicates is shown. E Experimental design: freshly transduced CB EG CD34⁺ cells were cultured for 21 days with TPO/FLT3L/EPO and without IL3/SCF. At that time point, IL3 and SCF were added or not to the culture medium and cells were grown for 14 more days (upper panel). Lower panel shows the mean ± SEM cumulative number of GFP⁺ cells of 3 biological replicates. F Experimental design: FL and CB EG cells were deprived of IL3 and/or SCF during 5 days of culture, after transduction or after their in vitro selection. The number of cells was counted at day 5 of culture ± cytokines. G Absolute numbers of EG GFP⁺ cells generated from transduced cells after transduction (condition with TPO/FLT3L/EPO). Mean ± SEM of 3 biological replicates. H Absolute numbers of EG GFP⁺ cells from in vitro selected EG cells. Mean ± SEM of 3 biological replicates. Statistical significance is indicated as p values (B,D,G,H: Kruskal–Wallis test, E: Mann–Whitney test). *: p < 0.05, **: p < 0.01