Research Paper Advance Articles pp 24394—24423

E3 ligase FBXW7 restricts M2-like tumor-associated macrophage polarization by targeting c-Myc

Figure 6. The role of FBXW7 in regulating M2 macrophage polarization is dependent on c-Myc. (A, B) Peritoneal macrophages (A) and BMDMs (B) derived from FBXW7f/f and Lysm+FBXW7f/f mice were stimulated with conditioned medium for the indicated time periods. The phosphorylated or total proteins related to M2 macrophage polarization were analyzed by immunoblotting. (C) Immunoblotting was used to analyze the interference efficiency of c-Myc siRNA in primary macrophages following stimulation with conditioned medium for 12 hours. (D, E) Immunoblotting (D) and qRT-PCR analysis (E) of Arg1 and Ym1 expression in primary macrophages from FBXW7f/f and Lysm+FBXW7f/f mice transfected with or without c-Myc siRNA and stimulated with conditioned medium. (F, G) Statistical analysis (F) and flow cytometry analysis (G) of the percentage of M2 macrophages (CD206+) in wild-type and FBXW7-knockout macrophages transfected with or without c-Myc siRNA and stimulated with conditioned medium. Data are shown as the mean ± SD and are representative of three independent experiments. ****P < 0.0001; n.s, no significance (two-way ANOVA (E, F)).