Fig. 7

FFAR2 inhibitor enhances the effect of anti-PD1 therapy. A–E WT mice were injected subcutaneously with LLC cells (1 × 10⁶ cells/mouse) and treated with FFAR2 inhibitor (GLPG0974, 5 mg kg⁻¹ per day; single esophageal gavage), intraperitoneally anti-PD1 (200 μg/mouse; every 4 days), FFAR2 inhibitor (GLPG0974) + anti-PD1 and PBS as control. Therapy was started at day 4 after tumor inoculation. LLC tumor growth (A) (n = 9) and tumor weight (B) (n = 9) were recorded. C and D Individual tumor growth of subcutaneous LLC tumors were recorded, and the number of survival tumor-bearing mouse at day 31 was shown (C) (n = 10), and overall survival of LLC tumor-bearing mice was recorded (Log-rank (Mantel-Cox) test) (D) (n = 10). E and F Representative gating strategy (E) and the percentage (F) of tumor-infiltrating CD4⁺ or CD8⁺-T cell of total CD45⁺ tumor-infiltrating leukocytes (TILs) in LLC tumors on day 30 post implantation were analyzed by flow cytometry (n = 5). G–I, Representative images of multicolor immunofluorescence staining for CD4 and CD8 in mouse LLC tumors (G). CD4⁺ T cells (H) (n = 5), and CD8 T cells (I) (n = 5) were quantified. J and K, Representative images of immunofluorescence staining for Arg1 in mouse LLC tumors (J). Arg1⁺ cells were quantified (K) (n = 5). A, B, F, H, I and K are shown as mean ± SEM, and the experiment was performed three times and a representative example is shown. A, B, F, H, I and K were analyzed by one-way ANOVA. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and NS, not significant)