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. Author manuscript; available in PMC: 2013 Mar 13.
Published in final edited form as: Cancer Res. 2011 Dec 15;72(3):666–675. doi: 10.1158/0008-5472.CAN-11-2774

Figure 4.

Figure 4

XPC Inhibits the Expression of Casp-2S at Both Protein and mRNA Levels. A, The expression of casp-2S and casp-2L in XP-C and XP-C+XPC cells were detected using Western blotting. B,C, The total RNA was isolated from XP-C and XP-C+XPC cells (B) or HCT116(p53−/−) cells transfected with either control or XPC siRNA (C), the transcript levels of casp-2S and casp-2L were determined using quantitative RT-PCR. N = 3, bars: SD, *: p< 0.05, compared with XP-C cells (B), **: p< 0.01 compared with control siRNA transfected cells (C). D, XP-C cells were transfected with different amounts of XPC containing pXPC3 constructs for 48 h. Whole cell lysates were prepared and the expression of XPC, casp-2S, casp-2L and Tubulin were analyzed using Western blotting. The intensity of each band was scanned; the relative amounts of casp-2 normalized to Tubulin were plotted. E, HCT116(p53−/−) cells were transfected with different amounts of XPC siRNA for 48 h. Casp-2S and -2L were detected as described in (D). F, Casp-2S promoter-luciferase construct (Del4) was transfected together with pGL4.73 plasmid into XP-C and XP-C+XPC cells for 48 h. The cultures were lysed and subjected to luciferase assays. N = 3, bars: SD, **: p< 0.01, compared with XP-C cells. G, HCT116(p53−/−) cells were transfected with control or XPC siRNA for 24 h. Casp-2S promoter activity was detected as (F). N = 3, bars: SD, **: p< 0.01, compared with cells transfected with control siRNA. H, ChIP analysis was performed with anti-XPC antibody and normal rabbit IgG in HCT116(p53−/−) cells. Four promoter sequences on the casp-2S promoter were analyzed by PCR.