Extended Data Fig. 4: 5-HT₇ is expressed on brain T_reg cells and required for the function of brain T_reg cells.

a, Scatter plot of T_reg signature genes (brain T_reg versus spleen T_reg cells). b, Heat map of the microarray analysis of T_reg cells from the brain, VAT, injured muscle, colon, pancreas and spleen. c, KEGG pathway analysis of T_reg versus T_conv cells, and brain T_reg versus spleen T_reg cells. d, Heat map of nervous-system-related genes of T_reg cells in the brain, VAT, injured muscle, colon (RORC⁺ and RORC⁻), lymph nodes and spleen. e, Scatter plot of nervous-system-related genes (brain T_reg versus spleen T_reg cells). f, Expression levels of Htr7 in the day-14 post-ischaemic or sham brains of diphtheria-toxin-treated DEREG or wild-type littermates, and brain T cells from wild-type ischaemic mice or spleen T cells from wild-type untreated mice. The gene expression levels were determined by microarray signals. Data were obtained from a single experiment of microarray, and confirmed by qPCR experiments. g–i, T_reg cells (5 × 10⁵) from spleens and lymph nodes from intact mice were transduced by CRIPSR–Cas9 (Htr7 gRNA or control gRNA) and transferred into Cd3e^−/− mice on day 5 after stroke onset. Flow cytometric analysis of 5-HT₇ (g), the number of T_reg cells (h) in the ischaemic brains of Cd3e^−/− mice, and neurological scores (i) on day 14 after stroke onset (n = 4 for control, n = 5 for 5-HT₇ knockout (KO)). Data are representative of two independent experiments. j, Flow cytometric analysis of 5-HT₇ in the ischaemic brains of wild-type mice on day 14 after stroke onset. Data are representative of three independent experiments. k, The relative expression levels of Htr7 in the ischaemic brains at day 14 after stroke onset were evaluated by qPCR. Brain T_reg cells were pooled from 6 mice, other data were n = 5 per group. Data are representative of two independent experiments and shown as mean ± s.e.m. P values were determined by two-tailed Student’s t-test.

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