Extended Data Figure 1: Analysis of one-month-old Csf1r^MeriCreMer;Braf^LSL-V600E;Rosa26 ^LSL-YFP mice.

a, The percentage of mice born from the cross depicted in Fig. 1a according to their genotype (n = 42), but without injection of hydroxy-tamoxifen (4-OHT) to test for adverse effects of 4-OHT administration. b, Flow cytometry analysis of YFP expression in blood leukocytes. Representative of n = 8 mice per genotype. c, Flow cytometry analysis of YFP⁺ cells in the liver. YFP⁺ cells, present only in Csf1r ^MeriCreMer+ (Cre⁺) mice (top), were gated as F4/80⁺CD11b⁺ Kupffer cells (bottom). Representative of n = 8 mice per genotype. d, YFP expression by immunofluorescence in the liver of Braf ^VE and Braf ^WT mice. YFP⁺ cells are F4/80⁺ Kupffer cells. Representative of n = 6 mice per genotype. Scale bars, 200 μm and 5 μm (insets). e, Total tissue-resident macrophage cell numbers per gram (g) of tissue were analysed by flow cytometry in Braf ^VE mice (n = 4) and Braf ^WT (n = 6). Circles represent individual mice. Unpaired two-tailed t-test. f, In situ analysis of phospho-histone H3 (pHis3) staining in YFP⁺ cells from brains of Braf ^VE and Braf ^WT mice. Circles represent individual mice (n = 3). Unpaired two-tailed t-test. g, RNA-seq analysis, heatmap of MAPK target genes in YFP⁺ microglia from Braf ^VE (n = 3) and Braf ^WT (n = 2) mice, values are displayed as z scores. h, Histological analysis of liver, lung, kidney and spleen in Braf ^VE and Braf ^WT mice. HE, haematoxylin and eosin. Representative of n = 4 mice per genotype. Scale bars, 200 μm and 10 μm (insets).

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