Extended Data Fig. 10: Related to Fig. 6. Id3-expressing macrophages control tumour growth.
From: The nuclear factor ID3 endows macrophages with a potent anti-tumour activity
a- Engulfment of KPC-1-memtdT cells (Casp.3/7 not cleaved) by BMDM expressing a control lentivirus (n = 7) in the presence or absence of α-SIRPA blocking antibody (n = 11), or an ID3 lentivirus (n = 18). Left: time from stable interaction between macrophages and tumour cells to tumour cells engulfment. Right: time from stable interaction between macrophages and tumour cells and the detection of Casp.3/7 cleavage, dots represent individual macrophages. b- Production of TNF by Human CD8+ T cells stimulated with anti CD3/CD28 activation beads, with supernatant from hiPSC-mac expressing lenti-control or lenti-hId3, and cocultured with Panc1 cells for 48 h or not for 3 days. Human CD8+ T cells are treated with cocktail of PMA, ionomycin, brefeldin A and monensin for 6 h, and TNF production measured by flow cytometry. n = 3 per group. c- (left) Numbers of NKT cells, γδT cells, CD19+ B cells, and CD4+ T cells in the tumours from 8–12 weeks-old C57BL/6 J mice two weeks after subcutaneous injection of 1 × 106 B16F10-luci-tdT cells into left and right flank, followed by intra-tumour injection of 5 × 105 BMDM expressing lenti-control (left flank) or lenti-mId3 (right flank) at day 7 post tumour injection. n = 20 mice per group from 3 experiments. (right) production of TNF by CD8+ T cells determined by flow cytometry. n = 5 mice/ group. d- Photoradiance analysis of liver tumour burden in 8–12 weeks-old C57BL/6j mice two weeks after subcutaneous injection of 1 × 106 B16F10-luci-tdT cells into left and right flank, followed by intra-tumour injection of 5 × 105 BMDM expressing lenti-control (n = 4), lenti-mId3 cells (n = 5) or not at day7 post tumour injection. Statistics: One-way ANOVA (a,b,d). unpaired two-tailed t-test (c). Dots represent individual mice (c, d). mean ± sd. ns, not significant. e- Schematic shows a hypothesis for the mechanisms that underly Id3-dependent anti-tumour activity of macrophages.
