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. Author manuscript; available in PMC: 2014 Oct 3.
Published in final edited form as: J Immunol. 2012 Aug 27;189(7):3439–3448. doi: 10.4049/jimmunol.1201168

Figure 5. Myeloid SOCS3 Influences Phagocytosis and Th1-Th17 Differentiation.

Figure 5

(A). Macrophages from SOCS3fl/fl and LysMCre-SOCS3fl/fl mice were polarized to the M1 phenotype with LPS for 48 h, and phagocytosis was assessed using the pHrodo™ E. coli BioParticles® Phagocytosis Kit for Flow Cytometry. *p<0.05. (B-E). LPS plus IFN-γ polarized M1 macrophages were used as antigen-presenting cells and cultured with naive CD4+ T cells isolated from the spleen of OVA-TCR transgenic OTII mice at a 1:5 ratio for Th1 and Th17 cell differentiation. Th1 cells were differentiated with IL-12 (10 ng/ml), anti-IL-4 (10 μg/ml) and OVA peptide (5 μg/ml); and Th17 cells were differentiated with TGF-β (5 ng/ml), IL-6 (20 ng/ml), IL-23 (10 ng/ml), anti-IFN-γ (10 μg/ml), anti-IL-4 (10 μg/ml) and OVA peptide (5 μg/ml). At day 4, cells were stimulated with PMA/Ionomycin plus GolgiStop for 4 h, stained for the surface marker CD4 and by intracellular flow for IFN-γ protein (B)., for IFN-γ and T-bet mRNA expression by qRT-PCR (C)., by intracellular flow for IL-17A protein (D)., or for IL-17A and RORγt mRNA expression by qRT-PCR (E). *p<0.05. Represents three independent experiments.