Fig. 2: LTβR regulates thymic tuft cell development.

a Intracellular staining of mTEC^low from control Foxn1^Cre and LTβR^TEC mice for expression of the tuft cell marker DCLK1. Bar graphs indicate absolute cell numbers and percentages within mTEC^low in Foxn1^Cre mice (closed symbols) and LTβR^TEC mice (open symbols) n = 7 biologically independent samples, over three independent experiments. Significant P values using two-tailed unpaired t test as follows: no. of tuft cells p = 0.0001, % tuft cells p = 0.002. b Confocal image of thymus section from Foxn1^Cre and LTβR^TEC mice, stained for the mTEC marker ERTR5 (magenta) and DCLK1 (cyan), representative of n = 3 independent biological samples. Scale bars denote 20 μm. c qPCR analysis of CD104⁻ mTEC^low FACS sorted from Foxn1^Cre (closed symbols) and LTβR^TEC (open symbols) mice, for expression of the tuft cell markers Pou2f3, Dclk1 and Trpm5, data are representative of three biological sorts. d Alymphoid 2dGuo-treated FTOC cultured for 4 days in the presence or absence of agonistic anti-LTβR (2 μg/ml) were analysed by flow cytometry for intracellular expression of DCLK1 to detect tuft cells. Tuft cells in freshly isolated adult WT mTEC^low are shown for comparison, data are representative of three independent experiments. e qPCR analysis of control (closed circles) and anti-LTβR stimulated (open circles) dGuo FTOC for expression of Pou2f3 and Dclk1, with levels of mRNA expression in adult mTEC^low (closed squares) for comparison, data are representative of three biological sorts from six FTOC cultured lobes/sort. f qPCR analysis of control (closed cirlces) and anti-LTβR stimulated (open circles) dGuo FTOC for expression of Ccl21a. All data are represented as mean ± SEM. ***P < 0.001 and **P < 0.01. Source data are provided as a Source Data file.