Extended Data Fig. 9. Tumour infiltrating leukocyte FACS and scRNA-seq analysis following treatment with anti-PD-L1, anti-TIGIT, and anti-CSF-1R.
a, Percentage of tumour macrophages (left) and representative FACS plots of tumour CD11b+ cell expression of F4/80 and CD86 following treatment (right). Data were collected at day 7 after treatment, and are representative of two independent experiments with n = 5 mice in each group. Left, data are mean +/ − SEM with one-way ANOVA with Tukey’s multiple comparisons. b, Growth of CT26 tumours in syngeneic BALB/c mice treated with anti-gp120 (left), anti-PD-L1 + anti-TIGIT IgG2a (middle), and anti-PD-L1 + anti-TIGIT mIgG2a + anti-CSF-1R (right). Data are representative of two experiments with n = 10 mice in each group. c, UMAP of tumour-infiltrating lymphocytes (top, n = 21, 575) and myeloid (bottom, n = 3, 734) cells coloured by cell types. d, Bubble plots showing marker gene expression for T and NK cells (left) and myeloid cells (right) as shown in (c). e, Volcano plots showing the gene expression of anti-PD-L1 + anti-TIGIT IgG2a versus control IgG2a (left), anti-PD-L1 + anti-TIGIT IgG2a + anti-CSF-1R versus control IgG2a (middle), and anti-PD-L1 + anti-TIGIT IgG2a + anti-CSF-1R versus anti-PD-L1 + anti-TIGIT IgG2a (right) in tumour macrophage and monocytes combined. f, g, Volcano plots showing the gene expression of anti-PD-L1 + anti-TIGIT IgG2a + anti-CSF-1R versus anti-PD-L1 + anti-TIGIT IgG2a in tumour CD8 + T cells combined (f) and CD4 Tregs (g). c-g, Single cell RNA-seq was performed on intratumoural CD45+ cells isolated from tumours at day 3 after treatment, and data are from one independent experiment with n = 5 mice in each group. In volcano plots, the broken y-axis was used to make the y-axis range comparable and for better comparison between treatments; P values were calculated by two-tailed Wilcoxon rank-sum test.