Fig. 3
C-myc regulates the induction of T-bet in unconventional T cells after TCR simulation. a Bar diagram shows C-myc mRNA expression (Mean ± SEM) of different thymocytes population from three 6–12-week-old TBGR mice performed by qPCR. Pre-gating is shown in Supplementary Fig. 1a. b C-myc mRNA expression for natural (TCRαβ+ or TCRγδ+ CD8αα+) and induced (TCRαβ+ CD8αβ+) IELs isolated from the small intestine of three 6–12-week-old TBGR mice and analyzed by qPCR. IELs were pre-gated for DAPI−CD45+CD4−. c Dot plot shows flow cytometric analysis of thymocyte subsets from 6- to 12-week-old C-mycΔ/ΔCd4 mice and littermate C-mycfl/fl mice as controls. Further flow cytometric analysis of DN thymocytes (d) and NK1.1− IELPs (e) as in c. c–e Bar diagrams represent percentages (left, Mean ± SEM) and absolute cells numbers (right, Mean ± SEM) of the respective thymocyte populations. f Histogram shows MFI of CD122 in thymic NK1.1−CD122+T-bet− IELPs as shown in e. g Flow cytometric analysis of BrdU incorporation in NK1.1− IELPs after 3 h of BrdU pulse labeling (contour plots) and bar diagram shows statistical analysis (Mean ± SEM) of BrdU+ subsets (right panel) of 2-week-old C-mycΔ/ΔCd4 mice and littermate C-mycfl/fl mice as controls for 5 mice per genotype. h Schematic model for the generation of bone marrow chimeras. i Representative dot plot shows flow cytometric analysis of different thymocyte populations from five OT-1 C-mycΔ/ΔCd4 bone marrow chimeras and C-mycfl/fll littermate controls. Cells were pre-gated on TCR Vα2 and numbers denote the percentage of cells in the gate (Mean ± SEM) of five mice per group. k Further flow cytometric analysis as in h for NK1.1− IELPs classified upon their CD122 and T-bet expression. j, l Bar diagrams show the percentage (Mean ± SEM) of the respective thymocyte subpopulation
