FIGURE 3.

SATB2 inhibits p63α-mediated perp activation. A, NIH 3T3 cells were transiently transfected with the indicated plasmids and quantitative real-time ChIP analysis was performed on the perp promoter (perp-1 site). Values were normalized to the amount of input DNA that was present. Real-time ChIP was also performed on mock-transfected cells as a control. B, a luciferase reporter assay using the murine perp promoter was performed in H1299 cells transfected with TA or ΔNp63α. A mutant reporter (MutD) with a point mutation in the perp promoter was used as a negative control. C, increasing amounts of T7-SATB2 were co-transfected with TAp63α into H1299 cells, and a perp luciferase reporter assay was performed. D, SCC9 cells were infected with lentivirus-expressing and shRNA targeting GFP or two different shRNAs directed against SATB2. Total RNA was extracted, and quantitative RT-PCR was performed using primers specific for perp. Values were normalized to gapdh housekeeper. IB, immunoblotting. E, SCC9 cells stably infected with lentivirus-expressing shGFP or shSATB2–2 were lysed, and Perp expression was analyzed by immunoblotting.