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Fig. 8. Phosphorylation represses the transactivating function of PPAR ${gamma}$ . (A) Western blot analysis of phosphorylation levels of PPAR ${gamma}$ in young and senescent 2BS and WI-38 cells. Total proteins were extracted, and western blotting was performed using specific antibodies against PPAR ${gamma}$ and phosphorylated PPAR ${gamma}$ (p-PPAR ${gamma}$ ) as indicated. Arrow indicates phosphorylated PPAR ${gamma}$ , and the observed doublet is reported as two translation initiation sites. (B) 2BS cells were transfected with expression plasmids pcDNA3.1 (–), pcDNA-PPAR ${gamma}$ (WT) or mutant pcDNA-PPAR ${gamma}$ (S84A) as indicated, together with wild-type p16-Luc. Cells were subsequently treated with 20 µM troglitazone or 10 µM pioglitazone. Luciferase activities were measured 24 hours after treatment. Values are the mean ± s.d. of triplicate points from a representative experiment (n=3), which was repeated three times with similar results. ^*P $<=$ 0.05.

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