Fig. 6. SD101 inhibits the generation of huMDSCs from PBMCs.

A Gating strategy to identify huMDSCs, its subtypes M- and G-MDSCs and M1 macrophages. PBMCs were treated with IL6 (20 ng/ml) + GMCSF (20 ng/ml) for 7 days in the presence or absence of 0.3 µM SD101. B Cells were treated with SD101 on D0, D2, and D7 and percentage of MDSCs (CD11b⁺CD33⁺HLA-DR^lo/−) was measured. C Ratio of M-/G-MDSCs was quantified; M-MDSCs: CD11b⁺CD14⁺CD15^-HLA-DR^lo/−, G-MDSCs:CD11b⁺CD14^-CD15⁺HLA-DR^lo/−. D Macrophage population was quantified: CD14⁺CD86⁺. E Annexin positive MDSCs were quantified. F PBMCs were treated once on D0 with SD101 (0.3 µM) for 48 h and MDSC was quantified. G PBMCs were stimulated with IL6 + GMCSF and treated with SD101 for 15 min and 4 h. FC analysis was performed to quantify pSTAT3 MFI in the MDSC gated cells and reported as fold change in the MFI of pSTAT3 positive cells. All the experiments were performed at least three times, and mean ± SEM was plotted in the graph.