Fig. 4

Expression of GPR43 in non-hematopoietic cells is necessary for GVHD-protective effect. WT B6 and Gpr43^−/− mice received BMT. a Total number of donor IFN-γ, TNF-α, and IL-17A positive T cells (n = 6 each). b Total numbers of CD4⁺CD25⁺Foxp3⁺ Treg cells, (n = 5 each, two-tailed unpaired t-test). c Serum IFN-γ, TNF-α, IL-6, and IL-17A levels from allogeneic recipients on day 14 after BMT (n = 6 each). d BMDCs treated with LPS were analyzed for CD40, CD80, and CD86 populations (n = 4 each, two-tailed unpaired t-test). e TNFα, IL-6, and IL-1β production by BMDCs stimulated overnight with LPS (n = 6 each, two-tailed unpaired t-test). f T cells from WT B6 or BALB/c were cultured with irradiated BMDCs for 72 and 96 h and analyzed for proliferation (n = 5 each, two-tailed unpaired t-test). g Total number of CD11b⁺ F4/80⁺ and CD11c⁺ 14 days after BMT (n = 4 each, two-tailed unpaired t test). h T cells from WT B6 incubated with anti-CD3 (2 μg per ml) and anti-CD28 (1 μg per ml) antibodies. Gene expression in 24, 48, 72, and 96 h after stimulation (n = 3 each). i Proliferation of stimulated splenic T cells for 72 h (n = 5 each, two-tailed unpaired t-test). j Irradiated splenocytes from BALB/c mice were co-cultured with effector T cells and Treg cells at different ratios and analyzed for T-cell proliferation (n = 3 each). k, l WT BALB/c mice received BMT. Survival and clinical GVHD score after BMT (n = 5 syngeneic and WT allogeneic each, n = 6 Gpr43^−/− allogeneic, log-rank test for survival, two-tailed Mann–Whitney U test for GVHD Score). m, n Chimeric [B6 → B6 Ly5.2], [Gpr4343^−/− → B6 Ly5.2], and [B6 Ly5.2 → Gpr43^−/−] animals received BMT. Survival and clinical GVHD score after BMT (n = 3 syngeneic each, n = 8 allogeneic each, log-rank test for survival, two-tailed Mann–Whitney U test for GVHD Score). o Serum LPS levels at day 14 after BMT (n = 4 naive, allogeneic, WT syngeneic, n = 10 Gpr43^−/− syngeneic, two-tailed unpaired t-test). p Serum FITC-dextran levels at day 14 after BMT (n = 4 each, two-tailed unpaired t-test). *P < 0.05, **P < 0.01, ***P < 0.001, error bars show the mean ± s.e.m. Data are representative of two or three experiments