Fig. 5

RASON knockout promotes macrophage phagocytosis of lung cancer cell lines. A-E, Effects of RASON KO on macrophage infiltration in LLC lung colonization tumors. A, Schematic representation of the FACS assay. Cells isolated from LLC lung colonization tumors were labelled antibodies against with antibodies against CD45, F4/80, CD11b, CD86, and CD206, and sorted using fluorescence-activated cell sorting (FACS). The percentage of activated macrophage cells (A-C), as well as M1 and M2 macrophages (E-F), was analyzed. CD45 + F480 + CD11b + cells represent activated macrophage cells, CD45 + F480 + CD11b + CD86 + CD11b- cells represent M1 macrophages, and CD45 + F480 + CD11b + CD86-CD11b + represent M2 macrophages. F-P, Effects of RASON KO on macrophage-mediated in vitro phagocytosis. (F) Schematic representation of the macrophage phagocytosis assay. Co-localization of macrophages with tumor cells in immunofluorescence staining and double-positive fluorescence signals in FACS analysis were used to indicate phagocytosis activity. (G-H) Representative immunofluorescence images of macrophage phagocytosis of tumor cells with or without RASON KO. (I-L) Representative percentage analysis of CFSE + CD14 + cells among the CFSE + cells (I, J) and mean fluorescence intensity of CD14 in CFSE + cells (K, L); (M-N) Quantitative analysis of G and H. (O-P), Quantitative analysis of I and J. Data are shown as mean ± S.D. and analyzed by Student’s t-test (C-E, M-P). * p < 0.05; ** p < 0.01; **** p < 0.0001