Figure 5.

Embelin impairs cytokine-induced MDSC generation and function in vitro.

Bone marrow (BM) cells were cultured in the presence of different cytokines for 4 days with or without embelin treatment. A. Accumulation of CD11b⁺Gr-1⁺ cells in BM cells is shown in contour plots and quantified. The percentage of CD124⁺ cells (B) or C/EBPβ⁺ cells (C) in CD11b⁺Gr-1⁺ cells was determined by flow cytometry. Contour plots are from one of four independent experiments, and data are expressed as mean ± SD. BM cells were cultured for 4 days with GM-CSF and G-CSF in the presence or absence of embelin, CD11b⁺ BM-derived MDSCs were enriched, and their immunosuppressive activity was determined. D. BM-derived MDSCs were co-cultured with CFSE-labeled T cells from naïve mice and stimulated with anti-CD3/CD28 antibodies. After 4 days, the percentage of CD4⁺ and CD8⁺ proliferating T cells is shown. E. BM-derived MDSCs were co-cultured for 4 days with CFSE-labeled splenocytes from OT-II transgenic mice and stimulated with OVA_323–339. The percentage of CD4⁺ proliferating T cells is shown. All results were obtained from three independent experiments *P < 0.05, **P < 0.01, ***P < 0.001.