Fig 3

PepO reprograms TAMs toward tumoricidal M1 in vivo. A Schematic of the TNBC model and PepO/PBS treatment regimen. 24 h after the second injection of PepO or PBS, mice were sacrificed, and the tumor tissue were collected (n = 3). B Volcano plot of DEGs in tumor tissue treated by PBS or PepO. The red and green dots represent significantly up-regulated and down-regulated genes, respectively. C Heatmap of key DEGs in groups including PBS control and PepO treated. D IHC staining of F4/80, iNOS and Arg-1 analysis and quantification of the positive cells in all field (n = 3, scale bar = 20 μm). E FACS dot plots showing % M2 macrophages (CD45⁺F4/80⁺CD11b⁺CD206⁺) and % M1 macrophages (CD45⁺F4/80⁺CD11b⁺CD86⁺ or CD45⁺F4/80⁺CD11b⁺iNOS⁺) derived from tumor tissue of TNBC bearing mice treated by PepO or not (n = 3). F The pro-inflammatory cytokine IL-6, and anti-inflammatory cytokine IL-10 in tumor tissue were detected using ELISA kit (n = 3). G Schematic of the TNBC model and the injection of clodronate/control liposomes through tail vein 24 h before treatments of PBS or PepO (n = 6). H, I Tumor growth profiles and tumor weight were detected in TNBC-bearing C57BL/6 mice treated with control liposomes + PBS, control liposomes + PepO, clodronate liposomes + PBS,clodronate liposomes + PepO, respectively (n = 6). J, K 1 × 10^5 PY8119 cells or an admixture of 1 × 10^5 PY8119 cells and 1 × 10^6 BMDMs treated as shown were injected into nude mice (n = 5 each). The tumors were collected at 4 weeks after injection and the tumor weights were compared. Two-tailed Student t test was used in (D, E, F). Two-way repeated-measures ANOVA with Sidak’s multiple comparisons test was used in (H). One-way ANOVA with Tukey’s multiple comparisons test was used in (I, K). Bar graphs represent mean ± SEM, *P < 0.05, **P < 0.01