Figure 5

Gene expression profiling of osteoblast and adipocyte markers in single cell-derived colonies. Total RNA was obtained at day 33 from each randomly selected colony as shown in Fig. 4. A, A representative agarose gel with amplimers from RT-PCR for adipsin, BSP, OCN, PPARγ, and C/EBPα. L32, internal control. Samples shown in lanes 1–3 and lanes 4–9 were collected from randomly selected ALP⁺ colonies in vehicle and BRL-treated cultures, respectively. One adipocyte colony (lane10) was also collected from a BRL-treated culture. B, Expression profiling of osteo/adipocyte marker mRNAs in ALP⁺/oil red O⁺ colonies in cultures treated with or without BRL. Data shown are from 9 ALP⁺/oil red O⁺ colonies; also shown for comparison are three ALP⁺ and three oil red O⁺ colonies. Data are representative of three independent experiments.