Figure 2 | Laboratory Investigation

Figure 2

From: Relationship between tumor-associated macrophage subsets and CD47 expression in squamous cell carcinoma of the head and neck in the tumor microenvironment

Figure 2

(a) CD47 surface expression in 5 SCCHN cell lines HSC-2, HSC-3, HSC-4, CA9-22, and Gun-1. All 5 cells were expressed CD47, and HSC-3 was the highest. (b) Phagocytosis of CFSE-labeled HSC-3 cells (green) by induced M2 macrophages stained with CD68 (pink) in a culture slide (arrow). Immunofluorescent staining confirmed that cancer cells were engulfed within 3 h by macrophages induced with our protocol. (c) Phagocytosis of HSC-3 cells by macrophages visualized by May–Grunwald–Giemsa staining. (i) A smear of HSC-3 cell suspension. HSC-3 cells had a small and a relatively reddish nucleus. (ii) A smear of peripheral blood. White blood cells including monocytes had a strong blue nucleus. (iii) Co-culture of induced macrophages and HSC-3 for 4 h. Large macrophages engulfed HSC-3 with small nuclei in vesicles in cytoplasm of the macrophages. (d) Phagocytosis of CD47+ HSC-3 by induced M1 (black bar) and M2 (white bar) subsets in vitro. A polyclonal sheep anti-human CD47-blocking IgG was added to media at concentrations of 0, 0.5, and 2 μg/ml at the time of the co-culture with macrophages and the CD47+ SCCHN cell line HSC-3. Summary experiments were repeated 8 times. Whole actual cell counts for analyses were 8162±695 of M1 and 7393±667 of M2.

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