Fig. 2. Loss of RORCE2 significantly decreases RORγt expression and Th17 cell numbers in vivo.

a The sequences of RORCE2 deleted with CRISPR/Cas9 technology (top). Allele knockout (KO) was confirmed by PCR (bottom left) and sequencing (bottom right). b Relative mRNA expression of RORγt gene in splenic CD4⁺ T cells from RORCE2^−/− (KO) or wild-type (WT) mice were measured by RT-qPCR assay. c, d Cells sorted from spleens of indicated mice were stained for CD4 and RORγt. The frequencies of CD4⁺RORγt⁺ Th17 cells in RORCE2^−/− and WT mice and mean fluorescence intensity (MFI) of RORγt in Th17 cells were analyzed by flow cytometry (c) and statistically evaluated (d). The numbers in quadrants indicate the percentage of cells in the quadrant. e Relative mRNA expression of IL-17A gene in splenic CD4⁺ T cells from indicated mice. f, g Frequencies of CD4⁺IL-17A⁺ Th17 cells in splenic CD4⁺ T cells from indicated mice. h, i RORγt⁺ Th17 frequencies in CD4⁺CD45⁺ Lin⁺ lymphocyte population from LP of small intestine of indicated mice. j, k MFI of RORγt in LP Th17 cells (j) and relative mRNA expressions of RORγt gene (k) in LP CD4⁺ T cells from the small intestine of indicated mice. l, m IL-17A⁺ Th17 frequencies in the CD4⁺CD45⁺ Lin⁺ lymphocyte population of LP from the small intestine of indicated mice. n Relative mRNA expressions of IL-17A gene in LP CD4⁺ T cells from small intestine of indicated mice. o–q Relative expression of RORγt gene in CD45⁺ Lin⁻CD127⁺ lymphocyte population (o) and frequencies of CD127⁺ RORγt⁺ ILC3 cells in the CD45⁺Lin⁻ lymphocyte population of the lamina propria from small intestine of indicated mice (p and q). Mean ± SEM are shown, n = 5 independent experiments, unpaired two-tailed Student’s t-test (b, d, e, g, i–k, m–o, q). Experimental mice were between 8 and 12 weeks of age, with no preference to gender and were maintained on a C57BL/6 background. Source data are provided as a Source Data file.