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. 2016 Apr 18;113(19):E2646–E2654. doi: 10.1073/pnas.1604268113

Fig. 3.

Fig. 3.

IFN-γ exposure inhibits antitumor antibody-dependent macrophage phagocytosis of B16F10 in vitro that can be rescued by combination PD-L1 and CD47 blockade. (A) Representative histograms of B16-F10 cell surface PD-L1, CD47, TRP-1, and CD200 expression before and after overnight treatment with 100 ng/mL IFN-γ as determined by flow cytometry. (B) Antibody-dependent phagocytosis of IFN-γ–treated (gray bars) or untreated (white bars) B16-F10 cells by bone marrow-derived C57BL/6J mouse macrophages treated with various combinations of anti–PD-L1 and/or tumor antigen-specific (αTRP-1) antibody with or without CD47 antagonist nanobody (A4). Phagocytosis is quantified as the percentage of F4/80-positive macrophages that have engulfed CFSE-positive B16-F10 cells as depicted in the representative FACS plots shown in C. The data shown are the mean (n = 3), and error bars indicate SD. ***P < 0.001; ****P < 0.0001 determined by one-way analysis of variance test in Prism.