Fig. 4

RvD1 prompts a PMN-dependent recruitment of anti-tumor monocytes in cancer models. a Percentage of classical (Ly6C^high) and non-classical (Ly6C^low) infiltrated monocyte subtypes in tumors from C57BL/6 mice subcutaneously transplanted with C3 cells and treated with vehicle or RvD1 (1 μg/kg) plus anti-IgG or anti-Ly6G, as reported in Fig. 3a. Data are expressed as percentage of myeloid cells as determined by FACS analysis using the gating strategy reported in panel (a). n = 4–6 mice tumors per group. *, P < 0.05; **, P < 0.01 (two-way ANOVA with Tukey’s multiple comparisons test). b Percentage of classical (Ly6C^high) infiltrated monocyte subtypes in tumors from C57BL/6 mice subcutaneously transplanted with TC-1 cells and treated with vehicle or RvD1(1 μg/kg) plus anti-IgG or anti-Ly6G as reported in Fig. 3a. Data are expressed as percentage of myeloid cells. n = 5–9 mice tumors per group. *, P < 0.05 (Mann Whitney test). c Number of human monocyte subsets transmigrated in the lower chamber of the Transwell insert 16 h after co-incubations with HeLa cells in the presence or not of PMN and treated with vehicle or RvD1 (100 nM). Number of monocytes are determined by measurements of CD14⁺CD16⁻ (classical) and CD14^lowCD16⁺ (non-classical) events recorded in 30 s of FACS acquisition. n = 4. *, P < 0.05 (two-way ANOVA and Tukey’s multiple comparison test). d Heatmap reporting the average concentration (pg/ml) of the indicated proteins in supernatants from PMN or PMN co-cultured with HeLa cells. e MCP-1 concentration in supernatants form HeLa/PMN coculture treated with vehicle or RvD1 (100 nM), as determined by ELISA. Data are expressed as fold change in MCP-1 concentration (pg/ml) compared to HeLa/PMN treated with vehicle. n = 6. *, P < 0.05 (Kolmogorov-Smirnov test). f Growth of HeLa cells during co-incubations with purified human CD14⁺monocytes exposed to vehicle or RvD1 (100 nM) and analyzed with an impedance-based real time cell analysis (ACEA). HeLa cells treated with vehicle or RvD1 in the absence of monocytes were used as control. n = 4. ***, P < 0.001 (HeLa Vehicle vs HeLa/classical monocytes Vehicle; HeLa RvD1 vs HeLa//classical monocytes RvD1; Wilcoxon matched-pairs signed rank test). g Correlation between the percentage of intratumor inflammatory monocytes and survival in mice transplanted with C3 cells. n = 26 XY pairs. Pearson r rank correlation was applied to analyze association between variables. Scattered plots with linear regression lines, Pearson r correlation coefficient, and two-tailed p value are reported