Research Paper Advance Articles pp 24394—24423

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

Figure 4. FBXW7 knockout facilitates M2 macrophage polarization in a tumor microenvironment-mimicking condition. (A) Peritoneal macrophages extracted from FBXW7f/f and Lysm+FBXW7f/f mice were treated with conditioned medium containing LLC supernatant. The mRNA expression of Arg1, Fizz1, and Ym1 were analyzed by qRT-PCR. (B) The protein levels of Arg1 and Ym1 in wild-type and FBXW7-knockout peritoneal macrophages were detected by immunoblotting after conditioned medium stimulation. (C, D) The mRNA (C) and protein (D) expression of Arg1, Fizz1, and Ym1 were analyzed by qRT-PCR and western blotting, respectively, in BMDMs incubated with conditioned medium. (E, F) Flow cytometry analysis (E) and statistical analysis (F) of the percentage of M2 macrophages (CD206+) in wild-type and FBXW7-knockout peritoneal macrophages stimulated with conditioned medium (n = 3 per group). (G, H) Flow cytometry analysis (G) and statistical analysis (H) of the percentage of M2 macrophages (CD206+) in wild-type and FBXW7-knockout BMDMs stimulated with conditioned medium (n = 3 per group). Data are shown as the mean ± SD and are representative of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (two-way ANOVA (A, C, F, H)).