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. 2017 Dec 4;114(51):E10981–E10990. doi: 10.1073/pnas.1712514114

Fig. 2.

Fig. 2.

Pretreatment of tumor epithelial cells with AZA and an HDACi leads to alterations in the numbers and activation of immune cell populations in tumor-associated ascites. ID8-VEGF-Defensin cells were pretreated (AZA, 500 nM; entinostat, 30 or 100 nM; givinostat, 100 nM) and injected into mice. Cells were analyzed from the drained ascites fluid (Fig. 1 A–C). (A) Immune cells per mL separated via Percoll gradient (n = 6 to 12 mice, two or three biological replicates). (B) CD45+ cells per mL identified via Percoll gradient and FACS (n = 6 to 11 mice, two biological replicates). Mean ± SEM is shown, and significances were determined by Mann–Whitney t test. (C–J) All cells from ascites were analyzed via FACS (n = 5 to 9 mice, one biological replicate). Mean ± SEM is shown, and significances were determined by one-way ANOVA. (C) CD45+ cells per mL of ascites. (D–J) Response of immune cell subpopulations to tumor cells pretreated ex vivo with AZA (A10). *P < 0.05, **P < 0.01, ***P < 0.001.