Fig. 1

ONO-4641 stimulates expansion of CD11b⁺Gr-1⁺ cells in naive mice. a, b Quantification of major lymphocyte populations, including B220⁺, CD3^-NK1.1⁺, CD3⁺γδT⁺, CD3⁺CD4⁺, and CD3⁺CD8⁺, in the lungs (a) and spleen (b) from mice intragastrically treated with vehicle or ONO-4641 (0.3 mg/kg/day) for 7 consecutive days (n = 4 in each group). Data are presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. c, d Representative two-parameter histograms (c) and quantification of CD11b⁺Gr-1⁺ cells in the lungs and spleen from mice intragastrically treated with vehicle or ONO-4641 (0.3 mg/kg/day) for 7 consecutive days. (n = 4 in each group). Data are presented as the mean ± SEM. *P < 0.05; **P < 0.01. e ONO-4641-expanded CD11b⁺Gr-1⁺ cells were divided into two populations: CD11b⁺Ly-6G−Ly-6C⁺ cells (monocytic myeloid-derived suppressor cell (MDSC)-like cells) and CD11b⁺Ly-6G⁺Ly-6C^low cells (granulocytic MDSC-like cells) (n = 3–4 in each group). f, g Representative cytospin images of CD11b⁺Ly-6G−Ly-6C⁺ cells (monocytic MDSC-like cells) (f) and CD11b⁺Ly-6G⁺Ly-6C^low cells (granulocytic MDSC-like cells) (n = 3–4 in each group) (g). h Representative two-parameter histograms of CD11b⁺Gr-1⁺ cells in the lungs and spleen from mice treated intragastrically or intraperitoneally with vehicle, C8-C1P (40 μg/kg/day), C16-C1P (50 μg/kg/day), SEW2871 (1 mg/kg/day), CYM-5442 (3 mg/kg/day), or ONO-4641 (0.3 mg/kg/day) for 7 consecutive days (n = 4 in each group). i Quantification of CD11b⁺Gr-1⁺ cells in the lungs and spleen from mice treated intragastrically or intraperitoneally with vehicle or the indicated molecules for 7 days (n = 4 in each group). Data are presented as the mean ± SEM. j S1pr1 and S1pr5 expression in CD11b⁺Gr-1⁺ cells sorted from the lungs of vehicle-treated or ONO-4641-treated mice (Day 7). (n = 3 in each group). Data are presented as the mean ± SEM. **P < 0.01; ***P < 0.001