Figure 3: PPARγ shRNA decreases PPARγ mRNA and protein levels and partially abrogates growth inhibition by Rosi. (a) 40 μg of nuclear extract protein from HTh74 cells transduced with either scrambled or PPARγ-specific shRNA (3 independent transductions) was separated on a 10% SDS-PAGE gel, transferred to PVD, and probed with PPARγ antibodies (sc-7196) as described in Figure 2(a). The blot was stripped and reprobed with antibodies to PPARδ (sc-7197). PARP was quantitated as a loading control. Shown above the protein blot are PPARγ RNA levels measured by qRT-PCR as in Figure 2(b). (b) Viable HTh74 cells either untransduced or transduced with scrambled (scr) or PPARγ shRNA were counted after 3 days treatment with 1 μM (white bars) and 10 μM Rosi (black bars). The DMSO vehicle represented by the dotted line is set to 100%. Shown is the % of vehicle control for each cell type (average of 3 different expts. ± SEM. *: significantly lower than vehicle, ; **: significantly lower than vehicle but higher than scr (10 μM), ).