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. 2016 Aug 9;149(3):320–328. doi: 10.1111/imm.12647

Figure 6.

Figure 6

Anti‐inflammatory and T helper type 2 (Th2) cytokine mRNA expression in CD4+ T cells co‐cultured with M1‐ and M2‐polarized peritoneal macrophages. (a) CD4+ T cells isolated from the spleen of wild‐type (WT) mice were stimulated with anti‐CD3/CD28 beads. These cells were co‐cultured with M1‐ or M2‐polarized peritoneal macrophages at a ratio of 1 : 10 (macrophages: CD4+ T cells). After co‐culturing, the floating cells were collected and stained for CD4 and F4/80 and analysed by flow cytometry. (b) Total RNA was isolated from CD4+ T cells co‐cultured with M1‐ and M2‐polarized peritoneal macrophages and then analysed by RT‐PCR to detect the T helper type 2 cytokines, Il4 and Il13, and the anti‐inflammatory cytokines, Tgfb and Il10. Cytokine mRNA expression levels were normalized to GAPDH. Data are representative of five independent experiments. Error bars represent SD. *P < 0·05 (Student's t‐test).