Skip to main content
. 2018 Apr 3;9(25):17443–17454. doi: 10.18632/oncotarget.24734

Figure 1. Conditional medium from NIH3T3/Src cells and colon cancer cells induced the polarization of macrophages.

Figure 1

(A) Ana-1 cells were treated with the supernatants from NIH3T3/p3.1, NIH3T3/Src cells or IL-4 (100 ng/ml) for 24 h or 48 h, respectively. The cells were analyzed with anti-F4/80 APC, anti-CD206 FITC and anti-CD163 PE using flow cytometry. Bars represent means ± SD (n = 3) for each treatment. *p < 0.05; **p < 0.01; ***p < 0.001. (B) Ana-1 macrophages were cultured in conditioned medium from NIH3T3/p3.1 or NIH3T3/Src cells for 48 h. Representative immunofluorescence images showed the expression and localization of F4/80 (red) and CD206 (green) in Ana-1 cells. DAPI is shown in blue. (Scale bar: 10 μm). (C) Ana-1 cells were cultured in the medium from NIH3T3/p3.1 or NIH3T3/Src for 48 h. The expression of iNOS and Arg-1 protein was analyzed by Western blotting. Intensity was quantified and normalized to β-actin. (D) Bone marrow-derived macrophages (BMDM) were cultured in medium from NIH3T3/p3.1, NIH3T3/Src, HCT116 or SW480 cells for 48 h. Cells were stained with anti- F4/80 APC, anti-CD206 FITC, then analyzed using flow cytometry. Bars represent means ± SD (n = 3) for each treatment. *p < 0.05; **p < 0.01; ***p < 0.001. (E) iNOS and Arg-1 expression in BDMD were analyzed by Western blotting. Intensity was quantified and normalized to β-actin.