Fig. 2

OCM/AS promotes M2-like TAMs polarization. (A) M0 MΦs (THP-1 MΦs, CD11b⁺CD14⁺) were treated with control medium or OCM and then, the expression of CD206 was analyzed by flow cytometry (n = 3). Representative flow cytometric plots (left) and summary bar chart (right). (B) M0 MФs (PBMC MΦs, CD11b⁺CD14⁺) were treated with OCM or ascites fluid (AS), and the expression of CD163 and CD206 was analyzed in OCM/AS treated macrophages or control macrophages by flow cytometry (n = 3). (C) The mRNA expression levels of CD163, IL10, TGFβ, CCL2, and NOS2 in M0 MФs (PBMC MΦs) treated with OCM/AS or control medium were measured by qPCR. Gene expression data were normalized to the reference gene 18 S, and the results are presented as the fold change relative to the control group. (D) M0 MФs (PBMC MΦs) were pre-stimulated with LPS/IFNγ (20 pg/mL, 20 ng/mL) to make M1 MΦs and were pre-stimulated with IL4/IL13 (20 ng/mL, 20 ng/mL) to make M2 MΦs. After that, MΦs (M1 & M2) were treated with OCM/AS or control medium, and then, the percentage of CD206, CD163, or CD86 were analyzed in macrophages and compared to M0 MΦs. The data were shown as the mean ± SEM and were analyzed by unpaired Student’s t-tests. n = 3, *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001