Extended Data Fig. 6: Related to Fig. 4. Id3-dependent cytokines and chemokines expression and lymphoid cells recruitment and activation in response to tumour.
From: The nuclear factor ID3 endows macrophages with a potent anti-tumour activity
a- SIRPA and Dectin-1 expression by flow cytometry of the main (Tim4+CD206+) and minor (Tim4+CD206hi) KCs subsets from the liver of Clec4fCreId3f/f mice and Id3f/f littermates, 2 weeks after intra-portal injection of 1 × 106 KPC-1-luci-gfp cells (KPC+), or from control mice without tumour (KPC−). n = 3 mice per group. b- Expression of Clec4n, Clec4d, Lrp1, and Siglec-G by RT-qPCR by KCs from the liver of Clec4fCreId3f/f mice and Id3f/f mice, n = 3 mice per group. c,d- Representative immunofluorescence micrograph of chemokines (c) and cytokines (d) expression by KCs of in the liver of Clec4fCreId3f/f mice and Id3f/f littermates 2 weeks after intra-portal injection of 1 × 106 KPC-1-gfp cells. n = 3 mice per group. e-g- Number of CD8+ T cells, Nkp46+NK cells, γδT cells, CD19+ B cells, CD4+ T cells, Ly6G+ granulocytes, Ly6Chi monocytes determined by flow cytometry in blood (e), and liver (f) and spleen (g) from 6-8 weeks old Id3f/f mice and Clec4fCreId3f/f mice (f,g), or from mice 2 weeks after intra-portal injection of 1 × 106 KPC-1-gfp cells (f). n numbers are indicated in green. h- Representative immunofluorescence micrograph for Lamp1, Nkp46, CD8, and GFP expression in liver sections from mice in (c). i- Expression of chemokines and cytokines by RT-qPCR by KCs from Clec4fCreId3f/f mice and Id3f/f littermates, cocultured with or without KPC cells for 48 h. n = 3 per group. j- IFN-γ production by flow cytometry by mouse splenic NK cells cultured for 3 days with supernatant from coculture in (h), n = 3 per group. Statistics: unpaired two-tailed t test (b,e,f,g), One-way ANOVA (a,i,j). Dots represent individual mice (a,b,e,f,g). mean ± sd. ns, not significant.
