Extended Data Fig. 7: Related to Fig. 5. SIRPA blockade and CD47 deficiency rescue Id3 deficient mice.
From: The nuclear factor ID3 endows macrophages with a potent anti-tumour activity
a- Dectin1 expression by flow cytometry (dots represent individual mice, n = 5/group), and CCL3, CCL4, and CCL5 expression by immunofluorescence (dots represent individual KCs) by Tim4+ KCs in liver from 8–12 weeks-old mice Clec4fCre Id3f/f mice and Id3f/f littermates, two weeks after intra-portal injection of 1 × 106 KPC-1-luciferease cells, treated with anti-SIRPA antibodies or IgG control antibodies. n numbers are indicated in green. b-d- Representative immunofluorescence micrograph of CCL3, CCL4, CCL5, by Tim4+ KCs at the boundary of GFP+ tumours in liver sections from mice in (a). e- Number of NKT cells, γδT cells, CD19+ B cells, CD4+ T cells per gram of tissue by flow cytometry analysis in mice from (a). n = 5 mice per group. f- Representative immunofluorescence micrograph of CD8, Lamp1, and Nkp46-expressing cells at the boundary of GFP+ tumours in liver sections from mice in (a). g- (left) CD47 expression by flow cytometry of tumour cell lines. (right) bioluminescence imaging and micrograph of liver tumour burden in C57BL/6 J mice 2 weeks after intra-portal injection of 1 × 106 KPC-1-Cd47+/+-luci-tdT cells or 1 × 106 KPC-1-Cd47−/−-luci-tdT cells, n = 7 mice per group from 2 independent experiments. h- Competitive in vivo proliferation assay between CD47−/− deficient and Cd47+/+ control KPC1 tumour cells in liver metastasis. 7 × 105 KPC-1-Cd47+/+-gfp tumour cells mixed with 7 × 105 KPC-1-Cd47+/+-tdT tumour cells or 7 × 105 KPC-1-Cd47−/−-tdT tumour cells in a 1:1 ratio are injected into c C57BL/6 J mice through intra-portal injection. Percentage of fluorescent cells expression in the liver and representative immunofluorescence of GFP and tdT performance after 2 weeks are shown. n = 5 mice per group. i- Photoradiance analysis of liver tumour burden in 8-12 weeks-old Clec4fCre Id3f/f mice and Id3f/f littermates two weeks after intra-portal injection of 1 × 106 KPC-1-luciferease-cd47+/+ cells or KPC-1-luciferease-cd47−/− cells, n = 4 mice per group. j- Numbers of CD8+ T cells, Nkp46+ NK cells, NKT cells, γδT cells, CD19+ B cells, CD4+ T cells per gram of tissue in the liver from mice in (i). n = 4 mice per group. Statistics: One-way ANOVA (a,e,i,j), Kruskal-Wallis test (a), unpaired two-tailed t-test (g,h). Dots represent individual mice (a,e,g,h,i,j). mean ± sd. ns, not significant.
