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. 2012 Nov 13;304(2):E197–E210. doi: 10.1152/ajpendo.00149.2012

Fig. 1.

Fig. 1.

Time-course of preadipocyte differentiation from rat retroperitoneal (RET) and subcutaneous (SC) adipose tissue in primary culture. Preadipocytes from RET (A) and SC (B) adipose tissues were plated in 35-mm Petri dishes at a density of 1.5 × 104/cm2. To initiate the process of differentiation, cells were cultured in DMEM-F12 medium-enriched differentiation cocktail as described in research design and methods. A and B: phase-contrast microscopy of SC and RET preadipocytes. Images were acquired at days 2, 4, 6, and 12 using a Leica microscope equipped with a ×20 objective. Squares in top represent areas selected for magnification shown in bottom. Undifferentiated cells, elongated fibroblast-like cells or more polygonal cells, still persisted in SC cultures. Scale bars, 60 μm for full-size images, 20 μm for magnifications. C: proteins were extracted before differentiation (day 0, D0) and after 2, 4, 6, 8, and 12 days in differentiation medium and processed for Western blotting using appropriate primary antibodies against PPARγ (1:1,000), aP2/FABP4 (1:2,000), hormone-sensitive lipase (HSL; 1:2,000), fatty acid synthase (FAS; 1:2,000), perilipin A, and β-actin antibody (1:8,000, 1:2,000). Data shown are representative results from 3 independent experiments.

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