Fig. 1. Identification of c-Rel-dependent monocytes by single-cell RNA sequencing.

a, Workflow for scRNA-seq of viable CD45.2⁺ cells isolated by FACS from LysM-cre × Rel^+/+ and LysM-cre × Rel^loxP/loxP mice harboring similarly sized B16F10 tumors (n=5 mice/group).

b, UMAP plots of 2830 intratumor myeloid cells that were selected for subsequent analyses. Each dot of the UMAP plot represents a single cell. Cell clusters are color-coded by associated cell types. c, Percentages of myeloid cell types inferred by Panel b.

d, Heatmap showing indicated gene expression across myeloid clusters.

e, Percentages of CD11b⁺CCR2⁺ cells in the indicated tissues of naïve and B16F10 tumor-bearing WT and Rel^−/− mice, as determined by flow cytometry; for naïve mice: WT, n=6 mice, Rel^−/−, n=7 mice; for tumor-bearing mice: WT, n=7 mice, Rel^−/−, n=8 mice.

f, Tumor growth in WT mice injected s.c. with B16F10 tumor cells plus CCR2⁺(rMo)/CCR2⁻ monocytes (n = 10 mice/group).

g, Percentages of CD44^high cells in intratumor CD8⁺ cells in f, as determined by flow cytometry. Statistical significance was determined by a two-tailed unpaired t-test. Data are presented as means ± s.e.m. (e-f), and pooled from two independent experiments (e) (*, P<0.05; **, P<0.01).