Fig. 10. CLA-induced IL-6 and IL-8 production predominates from nonadipocyte SV cells.

Cultures of SV cells containing newly differentiated human adipocytes were serum starved for 24 h and pretreated with 1 μg/ml brefeldin A for 1 h to prevent cytokine or chemokine secretion. Subsequently, cultures were treated with either a BSA vehicle or 30 μm trans-10, cis-12 CLA for an additional 12 h. Cultures were double stained for aP2 (rhodamine-conjugated anti-rabbit IgG = red) and subsequently for individual cytokines IL-6 or IL-8 (fluorescein isothiocyanate-conjugated anti-mouse IgG = green). Hoechst staining was conducted to identify nuclei positively. Fluorescent images (magnification, ×20) were captured as described under “Experimental Procedures.” Results shown are representative of two separate experiments from independent human subjects.