171765.fig.002a
(a)
171765.fig.002b
(b)
Figure 2: Protein and mRNA levels of PPARγ in thyroid cancer cells. (a) 60 μg of nuclear extract protein from various papillary and anaplastic thyroid cancer cells and 293 cells overexpressing PPARα, γ, or δ was separated on a 10% SDS-PAGE gel and transferred to a PVD membrane.The blot was blocked with 10% nonfat milk and incubated overnight with PPARγ rabbit polyclonal ab (sc-7196, Santa Cruz, CA) at a dilution of 1 : 500. Secondary antibodies were anti-rabbit IgG conjugated to horseradish peroxidase (GE Healthcare UK) at a 1 : 1000 dilution. PARP was quantitated as a loading control. (b) 200 ng of total RNA from the same thyroid cancer cells was subjected to qRT-PCR using an ABI PRISM7700 and the PPARγ-specific oligos and probe described in Section 2.5. PPARγ mRNA levels are shown per ng of 18 s rRNA.