Supplementary Figure 3.: Analysis of the expression and function of LILRB1 and LILRB2 in macrophages

(a) Flow cytometry-based measurement of the percentage of LILRB1 (blue) or LILRB2 (red) positive, non-immune cells from six samples of primary ascites fluid macrophages, and four samples of primary human colorectal carcinoma samples, as determined by fluorescence minus one controls. n.s. not significant, 2-way ANOVA with multiple comparisons correction. (b) Flow cytometry-based measurement of the percentage of LILRB1 (blue) or LILRB2 (red) positive tumor associated macrophages from four independent primary human colorectal carcinoma samples, as determined by fluorescence minus one controls. n.s. not significant, 2-way ANOVA with multiple comparisons correction. (c) Representative histogram plots from FACS analysis of LILRB1 and LILRB2 expression on primary human CD14+ peripheral blood monocytes (left), and day 7 ex vivo cultured macrophages derived from the same donor (right). Specific staining is indicated in blue; IgG control is indicated in red. (d) LILRB1 and LILRB2 expression levels, as measured by mean fluorescence intensity (MFI, left panel) or percent positive cells (right panel), in 4 pairs of primary monocytes (red) and ex vivo cultured macrophages from the same donors (blue). n.s., not significant. *** p < 0.001, 2-way ANOVA with multiple comparisons correction. (e) Flow cytometry-based measurement of phagocytosis by donor-derived macrophages of MHC I^– DLD1 (black) and MHC I⁺ DLD1 (red). Values are normalized to the highest level of phagocytosis observed in a given experimental replicate. Error bars represent s.d. of assays performed with n = 8 biological replicates. * p < 0.01, ** p < 0.001, 2-way ANOVA with multiple comparisons correction.