Fig. 2: Validation of cytokine receptor effects on macrophage polarization and identification of their essential downstream mediators.

a, Scheme of the validation experiment. Hoxb8FL control (tdTomato) cells were co-transferred with two different KO lines (GFP and blue fluorescent protein (BFP)) into the same immunized mouse before disease onset. b,d,f,h, Percentage of iNOS⁺ cell polarization in the single KO validation experiments compared to control, in the same animal (left) and scheme of the cytokine signaling pathway (right) for Ifngr1-KO (b), Tnfrsf1a-KO (d), Tgfbr1-KO (f) and Csf2ra-KO (h). The color code reflects the phenotype of M-iNOS polarization for the corresponding gene KO derived from the CRISPR screen shown in Fig. 1e–h. c,e,g,i, Percentage of Arg1⁺ cell polarization in the single KO validation experiments compared to control of the same animal (left) and scheme of the cytokine signaling pathway (right) for Ifngr1-KO (c), Tnfrsf1a-KO (e), Tgfbr1-KO (g) and Csf2ra-KO (i). The color code reflects the phenotype of M-Arg1 polarization for the corresponding gene KO derived from the CRISPR screen shown in Fig. 1e–h. b–i, n = 9 animals for Ifngr1-KO and Tgfbr1-KO transfer, n = 8 animals for Tnfrsf1a-KO and Csf2ra-KO transfer. Bar plots depict two-tailed paired t-test in b–h and a two-tailed paired Wilcoxon test in i. NS, P value > 0.05; *P value < 0.05, **P value < 0.01, ***P value < 0.001, ****P value < 0.0001. In b–i, asterisks indicate P values and FDR < 0.05 and absolute log₂(fold change) > 3 times the s.d. of noise distribution (Methods).

Source data