Fig. 1
NOTCH1 combined with other T-ALL oncogenes drives expansion of CB cells in vitro. a Experimental schema. Human CD34+ cord blood cells were transduced with GFP and Cherry lentiviral constructs, then plated on OP9-DL1 stromal feeders for varying periods of time prior to transplantation into immunodeficient NSG mice. b Flow cytometric tracking of doubly transduced (GFP+Cherry+, or G+C+) CB cells at each serial passage onto fresh OP9-DL1 feeders every 4–5 days. CB cells were transduced with lentiviral particles on days 0 and 5 only. Mean values ± range for duplicate experiments are plotted. c Flow cytometric tracking of NOTCH1ΔE-GFP, LTB-Cherry-transduced CB cells cultured as in (b). Gated live, hCD45+ events are depicted. Data are representative of multiple replicates. d Absolute growth of NOTCH1ΔE-GFP, LTB-Cherry doubly transduced cells (G+C+). Total cell number is extrapolated based on fold-increase in cell yield at each passage. Growth curves from three independent experimental replicates are shown. e Flow cytometric immunophenotyping of NOTCH1ΔE-GFP, LTB-Cherry doubly (G+C+) and nontransduced (G−C−) CB cell populations from days 19 and 33. Each plotted datapoint represents a separate culture. Mean ± SD values are plotted. *p < 0.05; **p < 0.01 (two-tailed t test with Holm−Sidak correction for multiple comparisons)
