Fig. 3

Lamina propria and intratumoural colon F4/80^hiCD11b⁺ macrophage subsets show differential CCR2-driven monocyte dependence. a Fluorescence-activated cell sorting histograms showing CCR2 expression profiles on distinct colon LP and intratumoural myeloid cell subpopulations: fraction I consists of MHCII^low and MHCII^hi cells, and fraction II consists of monocytes (P1) and monocyte-derived macrophages (P2 and P3). b Myeloid cell profiling in WT and Ccr2^−/− colon LP. Representative flow cytometry analysis with F4/80 and CD11b-expressing myeloid subpopulations, obtained from the colon LP of WT and Ccr2^−/− mice. Fractions I and II were further dissected for MHCII and Ly6C expression (left panel). The absolute numbers of eosinophils, F4/80^hiMHCII^hi and F4/80^hiMHCII^low tissue-resident macrophages and P1–P3 subpopulations are shown (right panel). The bar chart represents the mean number of mice in each group and the error bars represent the s.e.m. (WT; n = 7 and Ccr2^−/−; n = 14). c Myeloid cell profiling in WT and Ccr2^−/− colon tumours as described above for LP cells. Bar charts show the mean±s.e.m. of absolute numbers of eosinophils, F4/80^hiMHCII^hi and F4/80^hiMHCII^low tissue-resident macrophages and P1–P3 subpopulations obtained from Apc^Min/+ (n = 8) and Apc^Min/+Ccr2^−/− (n = 9) mice. Statistical significance was determined using an unpaired Student's t-test. **P<0.001; ***P < 0.0001; ns, not significant. Gating strategy is shown in Supplementary Fig. 4