Extended Data Fig. 7: sTNF-mediated TNFR1-specific signaling is dose-dependent and accelerates differentiation of human HSPCs into T cell precursors at the expense of their maintenance.
a, Scheme of assembly for the ATOs that were treated without (control) or with sTNF for 10 days. b, Absolute counts of CD45+ cells harvested from an ATO that was aggregated with a normalized amount of 7,500 cord blood-derived lin−CD34+CD38− HSPCs (n = 10 donors for all conditions except n = 3 for treatment with 0.25 ng/mL of sTNF). c-e, Flow cytometric analysis to identify CD7+CD5+ T cell precursors that were generated from the ATOs (c), of which some remained undifferentiated and expressed CD34 (d). Quantification of the impact of sTNF treatment on the cell counts of T cell precursors and undifferentiated CD34+ HSPCs compared to the control (n = 10 for all conditions except n = 3 for treatment with 0.25 ng/mL of sTNF) (e). f,g, Flow cytometric analysis to determine HLA-DR expression on CD1a-expressing T-lineage committed precursors (f) and quantification of the cellular fractions (n = 10 for all conditions except n = 3 for treatment with 0.25 ng/mL of sTNF) that were positive (g). Data are presented as mean±s.d. (b,e,g) and representatives (c,d,f) of three independent experiments. Data are analyzed by two-tailed mixed-effects analysis with Dunnett’s post-hoc test (b,g) and two-tailed mixed-effects analysis with linear trend test (e). n, biological replicates (b,e,g); exact P values are provided in the Source data.
