Fig. 6.

G9 suppresses tumor growth and cytokine expression and remodels the tumor immune microenvironment. (A) Estimated volume of KBP allografts treated with G9 or vehicle (n = 6 tumors per group). (B) Weight of KBP allografts treated with G9 or vehicle. (C) Immunoblot of N1^IC in tumor lysates of KBP allografts. Molecular weight markers are shown in kilodaltons. (D) Expression of N1^IC from C, determined by densitometry and presented relative to the loading control, β-actin. (E) Enzyme-linked immunosorbent assay of secreted IL-1β and CCL2 in conditioned media from overnight cultures of tumor cells from KBP tumors (n = 3 tumors per group). Cytokine levels are expressed per 10⁶ cells. (F–J) Quantification (by flow cytometry) of TAMs (Gr-1⁻ F4/80⁺ CD11b⁺), M2 TAMs (Gr-1⁻ F4/80⁺ CD11b⁺ CD206⁺) (F), cytotoxic T cells (CD8⁺) (G), helper T cells (CD4⁺) (H), regulatory T cells (CD4⁺ FoxP3⁺) (I), and myeloid-derived suppressor cells (Gr-1⁺) (J) in KBP allografts after 3 wk of treatment (n = 6 tumors per group). Values are expressed as percentage of CD45⁺ cells (F–H and J) or as percentage of CD4⁺ T cells (I). Error bars represent SEM. *P < 0.05 and **P < 0.01. P values are determined by two-way ANOVA (A) and Student’s t test (B and D–J). a.u = arbitrary units.