Effects of TNF-α, IL-6, and IL-1β on 3T3-L1 adipocyte differentiation

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Effects of TNF-α, IL-6, and IL-1β on 3T3-L1 adipocyte differentiation Effects of TNF-α, IL-6, and IL-1β on 3T3-L1 adipocyte differentiation. 3T3-L1 cells were differentiated for 4 days with thioglycolate-elicited peritoneal macrophages in direct coculture or in transwells. Effects of TNF-α, IL-6, and IL-1β on 3T3-L1 adipocyte differentiation. 3T3-L1 cells were differentiated for 4 days with thioglycolate-elicited peritoneal macrophages in direct coculture or in transwells. Cells were fixed and stained with Sudan IV dye as described in Materials and Methods. (A) Cells were imaged at ×200 magnification under a light microscope. “Mix” stands for direct coculture. (B) The lipid concentration was determined by measuring the absorbance at 550 nm. The number of PEC-C2D macrophages (Mφ) in each direct coculture is given at the bottom. Different letters indicate a significant difference. (C) 3T3-L1 cells were differentiated for 4 days and were then treated with TNF-α, IL-6, or IL-1β for an additional 4 days. High, medium, and low concentrations of the cytokines were as follows: 2, 1, and 0.5 ng/ml for TNF-α; 20, 10, and 1 ng/ml for IL-1β; and 10, 5, and 2 ng/ml for IL-6. At day 8, cells were fixed and stained with Sudan IV. Differentiation was determined by measuring the absorbance at 550 nm. The data are presented as means ± standard errors of the means for 3 to 6 independent replicates per treatment. An asterisk identifies a significant difference between the control and the treatment at each time point. A P value of <0.05 was considered significant. Linglin Xie et al. Clin. Vaccine Immunol. 2010; doi:10.1128/CVI.00494-09