Extended Data Fig. 5: Brain T_reg cells proliferate in both lymph nodes and brains.

a, Flow cytometric analysis of CD4⁺ T cells in cervical lymph nodes and brain from T_reg-cell-transferred Rag2^−/− mice. T_reg cells (2 × 10⁵ cells) from the cervical lymph nodes and spleens of ischaemic or sham-treated mice were transferred into Rag2^−/− mice on day 5 after stroke onset. The absolute numbers of CD4⁺ T cells in the brains of Rag2^−/− mice were measured on day 14 after stroke onset (n = 6). Data were pooled from two independent experiments. b, Kinetics of percentages of Ki67⁺ T_reg cells from brains and cervical lymph nodes of ischaemic mice at the indicated time points as determined by flow cytometric analysis of CD4⁺ T cells (n = 2 for days 0, 6, 11, 14 and 21; n = 3 for days 3 and 17). Data are representative of two independent experiments. c, Flow cytometric analysis of the frequency of T_reg cells among CD4⁺ T cells from the cervical lymph nodes of sham-treated and ischaemic mice on day 14 after stroke onset. The frequency of T_reg cells of CD4⁺ T cells was determined (n = 6 sham-treated mice, n = 12 ischaemic mice) Data were pooled from two independent experiments. d, The relative expression levels of Il2 in ischaemic brains at the indicated time points after stroke onset were evaluated by qPCR (n = 6 day 0, n = 3 others). Data are representative of two independent experiments. e, Flow cytometric analysis of IL-2⁺ cells from day 14 post-ischaemic brains and cervical lymph nodes gated as indicated. Data are representative of two independent experiments. f, Wild-type mice were injected with control IgG or anti-IL-2 antibody on days 8, 10 and 12 after stroke onset. Neurological scores were measured on day 14 (n = 8). Data are representative of two independent experiments and shown as mean ± s.e.m. P values determined by two-tailed Student’s t-test.

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