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. 2015 Nov 10;112(47):E6506–E6514. doi: 10.1073/pnas.1519623112

Fig. 3.

Fig. 3.

HAC–PD-1 shows enhanced tumor penetration and does not deplete peripheral T cells. (A) Representative fluorescence microscopy images of sectioned CT26 tumors deficient in PD-L1 (Bottom row) or transgenic for hPD-L1 (Top row) 4 h post-i.p. injection of anti–hPD-L1 AlexaFluor488 (green) and HAC AlexaFluor 594 (red). Nuclei (blue) were labeled with DAPI. (Scale bar, 500 μm.) (B) Representative flow cytometry of dissociated tumors from A showing HAC AlexaFluor 594 staining versus anti–hPD-L1 AlexaFluor 488 staining. Percentages are given in each positive quadrant. (C) Summary of flow cytometry studies from four PD-L1–deficient tumors and four hPD-L1 transgenic tumors. n.s., not significant. ***P < 0.0001, two-way ANOVA. Error bars represent s.e.m. (D) Relative abundance of peripheral CD4+ T cells (Left) or peripheral CD8+ T cells (Right) after 3 d of administration of vehicle (PBS), anti-mouse PD-L1, HACmb, anti-mouse CD47, or combinations of these agents to mice engrafted with CT26 tumors. Significance is indicated relative to the PBS control. ns, not significant. *P < 0.05, ***P < 0.001, one-way ANOVA.