Extended Data Fig. 5: Characterization of thymic tuft cells.
From: Thymic tuft cells promote an IL-4-enriched medulla and shape thymocyte development
a, DCLK1 and RFP (IL-25) in the thymus of C57BL/6 control and Flare25 mice. Confocal maximal projection. Scale bar, 5 μm. n = 3 mice, two independent experiments. b, RFP (IL-25) in (CD11c−CD45−EPCAM+) mTECs from C57BL/6 control and Flare25 mice. n = 5 mice; three independent experiments. c, Gating strategy for FACS sorting of CD11c−CD45−EpCAM+Ly51− mTECs from Flare25-reporter mice for qPCR analysis of mTEChigh (RFP−MHC-IIhigh), mTEClow (RFP+MHC-IIlow), and thymic tuft (RFP+) populations. d, qPCR analysis of expression of indicated genes of interest on populations sorted in c normalized to mTEChigh. Data are mean ± s.d. n = 3 mice. Two-way non-parametric ANOVA with multiple comparisons correction. e, Representative confocal maximum projection (10 μm) stained for KRT8/KRT18 (red) and DCLK1 (green). Scale bar, 50 μm. n = 3 mice, two independent experiments. f–i, Expression levels (normalized reads from Fig. 2b) from bulk RNA sequencing of small-intestinal (SI; n = 3 mice) and thymic tuft (n = 4 mice) cells. f, Expression of major MHC-I genes and B2m. FDR > 0.1. g, Expression of major MHC-II genes and Cd74 in small-intestinal tuft cells. h, Expression of minor MHC-II genes in thymic tuft cells. i, Expression of Tas2r family members in thymic tuft cells. g–i, Data are mean ± s.d. g, i, The red line corresponds to a cut-off of 5 reads per million; data for which the mean ± s.d. fall above this cut-off are indicated with an asterisk.
