Fig. 5: CLEC4A2 maintains macrophage identity and homeostasis by promoting CSF1- over CSF2-dependent differentiation in vitro.

a Principal component analysis (PCA) of the RNA-seq data from CSF1-BMDMs of WT and Clec4a2^−/− mice. b Heatmap of differentially expressed genes (P < 0.05) between WT and Clec4a2^−/− CSF1-BMDMs by RNA-seq (n = 3 mice, the heatmap represents one of two independent experiments). Differential expression was assessed using the two-sided Robinson and Smyth exact test with Bonferroni’s correction for multiple-hypothesis testing. c Expression of CD206 in WT and Clec4a2^−/− CSF1-BMDMs by flow cytometry (n = 5 mice each; the data are representative of two independent experiments). Two-tailed Student’s t-test. *P = 0.0158. d Schematic showing coculture of BM cells from 50% WT (CD45.1/CD45.2 heterozygous) and 50% Clec4a2^−/− (CD45.2) mice with CSF1. e Representative flow cytometry plots and proportion of each origin in CD11b⁺F4/80⁺CD206⁺ CSF1-BMDMs (n = 3 mice each; the data are representative of two independent experiments). Two-tailed Student’s t-test. *P = 0.0148. f Representative flow cytometry plots and percentages of 5-ethynyl-2′-deoxyuridine (EdU) incorporation in cocultured BMDMs (CD11b⁺F4/80⁺CD206⁺) with CSF1 (n = 3 mice each; one experiment). Two-tailed Student’s t-test. **P = 0.0048. g BM cells of 50% WT (CD45.1) and 50% Clec4a2^−/− (CD45.2) mice were cocultured with CSF2. Adherent cells were analysed by flow cytometry. Representative flow cytometry plots and proportion of each origin in CD11b⁺CD11c⁺F4/80⁺MHCII⁻ CSF2-BMDMs (n = 4 mice each; the data are representative of two independent experiments). Two-tailed Student’s t-test. **P = 0.0083. h Representative flow cytometry plots and percentages of EdU incorporation in cocultured BMDMs (CD11b⁺CD11c⁺F4/80⁺MHCII^-) with CSF2 (n = 3 mice each, one experiment). Two-tailed Student’s t-test. *P = 0.0208. All data are presented as mean ± SEM. Source data are provided as a Source Data file.