Fig. 3. Effects of the AhR on the differentiation potential of the clones. The cell clones were passaged to confluence and treated with the differentiation cocktail (DEX, IBMX and insulin). After 3 days, the cells were re-fed with fresh differentiation medium without DEX and IBMX, and maintained for several days. (A) On day 6, cells were fixed and stained with oil red O. The dishes in the middle lane show the cells expressing vector mRNA (V) (left and right) and wild-type 3T3-L1 cells (WT) (center). AS, cells expressing antisense mRNA; S, cells expressing AhR sense mRNA. (B) Induction of GPDH activity in the clones. Enzyme extracts were prepared at days 0 and 6 from the three independent clones. GPDH activity was measured as described in Materials and Methods. The results presented are the means from three separate determinations on three independent clones (n=9). (C) Expression of C/EBP, PPAR2 and aP2 mRNA in the clones. The cell clones were treated as described in the legend to Fig. 4. On days 0 and 6, total RNA was extracted and the expressions of C/EBP, PPAR2 and aP2 mRNA were determined by northern blotting. AS, cells expressing antisense mRNA; S, cells expressing AhR sense mRNA.