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. 2018 Jul 23;9:1699. doi: 10.3389/fimmu.2018.01699

Figure 5.

Figure 5

Reversible myeloid differentiation blockage of early-stage MDSCs (e-MDSCs) is interleukin-6 (IL-6) dependent. Bone marrow (BM) cells isolated from tumor-free BALB/c mice were cocultured with rIL-6, tumor cell supernatant, and tumor cells to induce e-MDSCs in vitro. And BM cells treated with GM-CSF (10 ng/mL) were regarded as normal control. (A) The percentage of CD11b+Gr-1− was detected by flow cytometry. (B) The percentage of F4/80+ and MHCII+ cells in CD11b+Gr-1− myeloid-derived suppressor cells (MDSCs) was detected using flow cytometry. BM cells were isolated from tumor-free mice and cultured with GM-CSF for 3 days with or without 4T1 cells. IL-6 receptor Ab was utilized to block IL-6 downstream signaling. (C) The percentage of CD11b+Gr-1−, CD11b+Gr-1+, F4/80+, and MHCII+ cells was detected using flow cytometry. (D) Primary CD11b+Gr-1−F4/80−MHCII− cells were isolated using the BD FACSAria™II cell sorter and induced with GM-CSF. The percentage of CD11b+Gr-1−, CD11b+Gr-1+, Ly6G (1A8)+ cells, F4/80+ cells, and MHCII+ cells was detected by flow cytometry (*P < 0.05; **P < 0.01; and ***P < 0.001).