Figure 4.

Immune suppression by MDSCs sorted by Gfi1:GFP expression. (a) Sorted MDSC subsets were cocultured with CFSE-labeled splenic CD4⁺ T cells isolated from naïve spleen by magnetic cell sorting. Anti-CD3 and anti-CD28 antibodies were applied to induce CD4⁺ T-cell proliferation. After 72 h, the proportion of proliferating CD4⁺ T cells was evaluated by CFSE staining and flow cytometry. Representative graphs show data from three independent replications. (b) Sorted CD11b⁺Gr-1⁺CD48⁻ G-MDSCs and CD11b⁺Gr-1⁺CD48⁺ M-MDSCs were analyzed by real-time qPCR. Probes detecting NOS2 and Arg-1 were used to quantify mRNA levels. Relative expression was normalized to bone marrow derived granulocytes from naïve mice. Data are mean ± SEM of 6–10 mice per group. The experiment was repeated three times, and the values presented are from one representative experiment. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001, ^∗∗∗∗P < 0.0001.