Extended Data Fig. 2: scRNA-seq in small and large size B16F10 tumours.

a, Schematic for harvesting B16F10 tumours at small (150 mm³) and large (300 mm³) volumes and processing for scRNA-seq. b, UMAP plot of cells from 4 pooled large B16F10 tumours (300 mm³) from each genotype. c, Dot plot displaying selected gene markers across all clusters. The colour intensity represents the average expression while the size of dots indicates the percentage of cells expressing each gene. d, Violin plots showing the expression of ATF4 across all clusters in small sized tumours and e, in large sized tumours. f, Bar plot displaying the normalized Log2 fold change of cell types in each cluster in small sized tumours and g, in large sized tumours. h, Tumour growth curves of Atf4^wt/wt and Atf4^Δ/Δ mice treated with Isotype or anti-CD8 antibody following 5×10⁵ B16F10 cell injection (n = 3–5 biologically independent samples per group). Values represent mean ± SEM. Two-way ANOVA analysis (until day 18). i, Violin plots showing the expression of the indicative markers at the CAFs cluster. j, Bar plot displaying the eight most significant Reactome pathways enriched in Atf4^wt/wt CAFs using the genes upregulated in Atf4^wt/wt. k, Violin plots showing the expression of the indicative CAF and melanoma markers at the CAFs subclusters and melanoma cells (small B16F10 tumours).

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