Figure 7. In vivo cytokine signaling is always required for SP8 cell generation.

(a) Profiles of TCRβ^hiCCR7⁺ thymocytes from the indicated mice are shown, and numbers in boxes within the profiles indicate frequency of SP8 cells among TCRβ^hi thymocytes (left). Bar graph displays SP8 thymocyte numbers in each mouse strain (right). Data are from 2-5 mice combined from 2-5 experiments. (b) Comparisons of SP8 thymocyte frequencies (left) and numbers (middle) in γc-sufficient (γc^WT) and γc-deficient (γc^cKO) Runx3d^YFP/YFP mice, and Runx1 and Cbfb mRNA abundance in SP8 cells from the same mice (right). Cell numbers and frequencies are from 8 mice combined from 5 experiments, quantitative PCR data are technical triplicates and one of two experiments is shown. (c) Comparison of Runx1 mRNA expression in electronically sorted SP8 thymocytes from Runx3d-sufficient (Runx3d^WT) and Runx3d-deficient (Runx3d^YFP/YFP) mice. Data are replicates from two representative experiments out of a total of four experiments done. (d) Runx1 and Runx3d mRNA expression in electronically sorted SP4 and SP8 thymocytes from B6 mice. Data are technical triplicates and one of two experiments is shown. (e) Profiles of MHC class II-selected TCRβ^hi thymocytes from γc-sufficient (γc^WT) and γc-deficient (γc^cKO) ThPOK^KOβ2m^KO mice are shown, and numbers in boxes within the profiles indicate frequency of SP8 cells among TCRβ^hi thymocytes (left). The bar graph compares the number of MHC class II-selected SP8 thymocytes in γc-sufficient and γc-deficient ThPOK^KOβ2m^KO mice (right). Data are from 3 mice combined from 3 experiments. Mean and s.e.m. are shown. *P < 0.05, **P < 0.01, ****P < 0.0001.