Extended Data Fig. 4: Raptor deficiency induces selective phenotypic changes in fate-mapped T_H17 cells.

a–g, WT and Rptor^Il17aCre (R26R^eYFP) mice were immunized with MOG, and YFP⁺ cells from dLN were analysed at day nine post-immunization. a, Frequency of YFP⁺ cells (n = 7 per genotype). b, Efficiency of Rptor deletion (left; n = 7 per genotype) and flow cytometry analysis of phosphorylated S6 (S235/236) and 4E-BP1 (T37/46) (right) in YFP⁺ cells. c, Flow cytometry analysis of active caspase-3 and 7-AAD staining in YFP⁺ cells. d, Flow cytometry analysis of CXCR3 and CCR6 expression on YFP⁺ cells. e, Cytokine production by YFP⁺ cells from dLN (n = 7 per genotype). f, Real-time PCR analysis of Tbx21 (n = 8 per genotype), Rorc (n = 6 per genotype), Il12rb2 (n = 7 per genotype) and Il23r (n = 7 per genotype) expression in YFP⁺ cells. g, Flow cytometry analysis of Foxp3 expression. h, i, Cytokine production (h, i) and proliferation (h) of YFP⁺ cells from dLN of the indicated mice after four days of stimulation with MOG alone (h) or with MOG plus IL-23 (i) (n = 7 per genotype). j, Sorted YFP⁺ cells were stimulated with IL-23 or IL-12 for 30 min in vitro and stained with specific antibodies to phosphorylated STAT3 (left), phosphorylated STAT4 (right), or isotype controls. Data are means ± s.e.m. and representative of seven (a), three (b–f, j), two (g), or five (h, i) independent experiments. Numbers in plots represent frequencies of cells in quadrants; numbers within histograms represent mean fluorescence intensities. Student’s t-test (two-sided) was used in b, and Mann–Whitney U-test (two-sided) in a, e, i, to determine statistical significance.

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