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. 2017 Oct 13;8:1300. doi: 10.3389/fimmu.2017.01300

Figure 3.

Figure 3

Figure 3

TEPP-46 reduces PD-L1 expression on tumor and immune cells in vivo in the CT26 colon carcinoma animal model. Single cell suspensions from CT26 tumors or dLN isolated on day 15 post-tumor induction were stained for flow cytometry (A–C). Representative FACS histograms of PD-L1 cell surface expression on CD45− or CD45+ cells from tumor dLN or tumors (A). Representative FACS histograms of PD-L1 cell surface expression on different CT26 infiltrating immune cell populations (B). (C) PD-L1 MFI for CD45− or CD45+ cells from naïve LN, dLN, or tumors (T). Mice were inoculated with CT26 tumors and injected i.p. with vehicle or TEPP-46 every 2–3 days, beginning on day −1 pre-tumor induction (D–I). Tumors and dLNs were isolated on day 15 post-tumor induction. MFI of PD-L1 cell surface expression on different immune populations in dLN (D) or tumors (E). Representative histogram of PD-L1 expression on CD45− cell from vehicle or TEPP-46-treated tumors (F). PD-L1 MFI for CD45− cells from tumors (G). Total pdl1 RNA expression in cells isolated from dLN [(H), left] or tumor [(H), right]. Statistics are performed as one-way ANOVA with Bonferroni post-test (C) and two-tailed unpaired t-test (D,E,G,H); *P < 0.05, **P < 0.01, and ***P < 0.001. (I) Expression of proglycolytic and Hif-1α-dependent genes (glut1, ldha, and phd3 top panels, sdha, hk2, and pkm2, bottom panels) in tumor biopsies isolated on day 15 post-tumor induction.