Panel A shows bar plots comparing myeloid cell populations in the blood, spleen, and bone marrow. The x-axis represents different cell populations, and the y-axis represents the percentage of CD45 positive cells. Panel B shows flow cytometry plots illustrating the gating strategy for identifying myeloid cell populations in EO771 tumors. Panel C shows bar plots comparing myeloid cell populations in WT and CD1d-knockout EO771 tumors at day 15 after tumor cell injection. The x-axis represents different cell populations, and the y-axis represents the percentage of CD45 positive cells. Panel D shows flow cytometry plots and a bar plot comparing the ratio of major histocompatibility complex class 2 high versus major histocompatibility complex class 2 low CD64 positive cells in WT and CD1d-knockout tumors. Panel E shows flow cytometry plots illustrating the gating strategy for identifying lymphoid cell populations in EO771 tumors. Panel F shows bar plots comparing lymphoid cell populations in WT and CD1d-knockout EO771 tumors at day 15 after tumor cell injection. The x-axis represents different cell populations, and the y-axis represents the percentage of CD45 positive cells. Panel G shows flow cytometry plots illustrating the gating strategy for identifying cell populations in mixed bone marrow chimeras. Panel H shows a line graph of EO771 tumor growth in mixed bone marrow chimeras. The x-axis represents days after orthotopic injection, and the y-axis represents tumor size in square centimeters. Panel I shows bar plots comparing cell populations in the spleen, bone marrow, and peritoneal cavity of mixed bone marrow chimeras. The x-axis represents different cell populations, and the y-axis represents the percentage of CD45 positive cells.
Immune infiltrates in EO771 tumors in WT and CD1d-KO mice. (A) Flow cytometry analyses showing frequencies for the depicted myeloid populations in the blood, spleen, and BM from WT and CD1d-KO mice in steady state (n = 2–5; data are pooled from two independent experiments). Data are shown as the mean ± SEM. (B) Flow cytometry plots showing gating strategy for myeloid cell populations from EO771 tumors. (C) Flow cytometry analyses of immune infiltrates in EO771 tumors (from WT and CD1d-KO mice) at day 15 after tumor cell injection, depicting frequencies of the indicated cell populations (n = 20; data are pooled from seven independent experiments). Data are shown as the mean ± SEM. (D) Flow cytometry plots showing gating strategy and ratio (top, right) of MHCII-hi versus MHCII-low CD64+ cells (CD45+CD11b+CD64+Ly6C−Ly6G−) in WT and CD1d-KO TMEs (n = 20; data are pooled from seven independent experiments). Data are shown as the mean ± SEM. (E) Flow cytometry plots showing gating strategy for lymphoid cell populations from EO771 tumors. (F) Flow cytometry analyses of immune infiltrates in EO771 tumors (from WT and CD1d-KO mice) at day 15 after tumor cell injection, depicting frequencies of the indicated cell populations (n = 4–14; data are pooled from three to five independent experiments). Data are shown as the mean ± SEM. (G and H) Mixed BM chimeras (WT:CD1d-KO; 50:50) were generated by cotransferring WT (CD45.1+) and CD1d-KO (CD45.2+) BM into irradiated recipients (CD45.1+CD45.2+). After reconstitution, mice were orthotopically injected with EO771 cells and immune infiltrates were analyzed at day 15. (G) Flow cytometry plots showing gating strategy for the depicted cell populations. (H) EO771 tumor growth in chimeras (n = 8; data are pooled from two independent experiments). (I) Flow cytometry analyses showing frequencies for the depicted cell populations in the spleen, BM, and peritoneal cavity from mixed BM chimeras (WT:CD1d-KO; 50:50) in steady state (no tumor injection, n = 5; data are pooled from two independent experiments). Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, unpaired t test.