TAMs directly contribute to tumor hypoxia.

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TAMs directly contribute to tumor hypoxia. TAMs directly contribute to tumor hypoxia. A, Immunostaining of LLC tumors grown in mice for TAM by using S100A8 (red) and hypoxia by using pimonidazole (PIMO; green) antibodies. Nuclei are shown in blue with DAPI counterstaining. B, FACS analysis demonstrating that CD11b and F4/80 double-positive TAMs are pimonidazole-positive. C, Gene expression in CD11b and F4/80 double-positive TAM isolated from LLC tumors compared with those in cultured BMDM. Data are the mean ± SEM from triplicate determinations. D, Two-photon microscopy images of the dorsal window chamber whereby 5 × HRE-GFP–expressing LLC tumors had been implanted. Images were taken at 24 hours after a single intratumoral injection of PBS (+PBS) or PBS containing FACS-sorted TAM (+TAM). Scale bars in A and D, 100 μm. E, Representative FACS plots demonstrating HoechstbrightKeratin+ as aerobic tumor cells (red boxes) and HoechstdimKeratin+ as hypoxic tumor cells in LLC tumors grown in mice treated with Veh or Clod. F, Quantification of aerobic or hypoxic tumor cells in E. Data are the mean ± SEM for n = 6 mice per group. *, P < 0.05 by Student t test. G, TCGA analysis between CD68 and HIF1A in 513 patients with adenocarcinoma NSCLC. P value is indicated in the graph. H, Growth of LLC tumor treated with Veh or Clod immediately prior to a single dose of 20 Gy ionizing irradiation. *, P < 0.05 by two-way ANOVA. Hoibin Jeong et al. Cancer Res 2019;79:795-806 ©2019 by American Association for Cancer Research