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. Author manuscript; available in PMC: 2019 Jul 1.
Published in final edited form as: J Immunol. 2018 May 11;201(1):264–277. doi: 10.4049/jimmunol.1800129

Figure 1. PD-L1 expression profiles on MDSCs in vitro and in vivo.

Figure 1

A. Bone marrow cells were cultured with GM-CSF for five days. Cells were stained with CD11b-, Gr1-, Ly6G- and Ly6C-specific mAbs. CD11b+Gr1+ cells were gated and plotted for PD-L1 levels (a). CD11b+ cells were further gated based on Ly6G and Ly6C levels. Ly6G+Ly6Clo and Ly6GLy6Chi cells were plotted for PD-L1 levels (b & c). Shown are representative results of one of three independent experiments. B. BM cells were cultured with AT3 tumor cell-conditioned medium for five days. PD-L1 levels in CD11b+Gr1+ (a), CD11b+Ly6G+Ly6Clo (b) and CD11b+Ly6GLy6Chi (c) cells were analyzed as shown in A. Shown are representative results of one of three independent experiments. C. Tumor tissues were collected from mammary carcinoma AT3 tumor-bearing mice and prepared for single cells. Cells were stained with CD11b-, Gr1-, Ly6G- and Ly6C-specific mAbs. PD-L1 levels in tumor-infiltrating CD11b+Gr1+ (a), CD11b+Ly6G+Ly6Clo (b), and CD11b+Ly6GLy6Chi (c) cells were analyzed as in A. Shown are representative results of one of three independent experiments. D. Tumor tissues were collected from colon carcinoma CT26 (left panel) and MC38 (right panel) tumor-bearing mice, and analyzed for PD-L1 levels in tumor-infiltrating CD11b+Gr1+ (a), CD11b+Ly6G+Ly6Clo (b), and CD11b+Ly6GLy6Chi (c) cells were as shown in A. Shown are representative results of one of three independent experiments. E. PD-L1 MFIs between CD11b+Ly6G+Ly6Clo and CD11b+Ly6GLy6Chi MDSCs from the above five in vitro and in vivo MDSC models were compared. Column: mean; Bar: SD. F. J774M cells were stained with CD11b-, Gr1-, and PD-L1-specific mAbs and the CD11b+Gr1+ cells were analyzed for PD-L1 level.