(A) IL-13Rα1-GFP reporter mice were given STAT6 inhibitor (STAT6 I) or diluent (No I) and 5 days later HR+ETPs were sorted and cultured (3 x 103 cells per well) on OP9-DL1 cells in the presence of IL-7+ Flt3L. Ten days later the cultures were assessed for commitment to the T-cell lineage by measuring CD25 expression on 7-AAD−CD45+CD11b− cells. (B) HR+ETPs were isolated from IL-13Rα1-GFP reporter mice, transduced with Tcf-1-RV or Empty RV and cultured (0.7 x 103 cells per well) on OP9-DL1 cells in the presence of IL-7+Flt3L for 10 days. Commitment to the T-cell lineage was assessed by measuring CD25 expression on 7-AAD−CD45+CD11b− cells.
In all contour plots the numbers indicate the percent of CD25+ cells and bar graphs show the number of CD25+ cells compiled from 3 independent experiments. Each bar represents the mean ± SD for each population. ***p<0.001 as determined by two-tailed, unpaired Student’s t-test.