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. 2010 Dec;12(12):969–979. doi: 10.1593/neo.101054

Figure 3.

Figure 3

Smad2 requires endogenously expressed IKKα for activation of the ΔNp63 promoter. (A) Luc reporter assay in A431 cells with IKKα silencing. Twenty-four hours after the siRNA (Csi, IKKαsi) introduction, cells were seeded onto 24-well plates to perform the luciferase assay with 2kΔN-luc and expression vectors of the Smad proteins and IKKα. Cells were incubated with (+) or without (-) TGF-β1 (TGF-b1) as described in the legend to Figure 2. Luciferase activity 48 hours after the 2kΔN-luc transfection was determined. Relative luciferase activity (RLA) is represented as mean ± SD; n ≥ 3). Statistic significance: **P <. 01, *P <. 05, versus control experiment with Vec. (B) IKKα silencing in FaDu cells. We determined luciferase activities produced by the Csi- and IKKαsi-transfected cells cultured with 10% FBS. Results with A431 under the same conditions are also shown. (C) Western blot analysis of A431 and FaDu cells with IKKα silencing. Day 3 cells after the transfection of Csi and IKKαsi were analyzed for IKKα (85 kDa) and β-actin (43 kDa). Relative amounts of IKKα were obtained by standardization with β-actin for each cell line.