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. 2011 Oct 20;286(50):43204–43213. doi: 10.1074/jbc.M111.257105

FIGURE 6.

FIGURE 6.

TAp63α increased levels of RelA-dependent genes. MCF10A cells were transfected with the TAp63α expression plasmid (0–1.5 μg). A and B, Ready-To-GlowTM Secreted Luciferase assay for the NF-κB/RelA response element-driven Metridia luciferase activity. 12 h after transfection of H1299 (A) and MCF10A (B) cells with the indicated constructs, the medium was replaced, and then after 16 h, samples of the medium were analyzed for secreted Metridia luciferase activity. The -fold induction was calculated following the substrate addition. C and D, luciferase reporter assay for the Cdkn1a (C) and Bbc3 (D) promoters. MCF10A cells were transfected with the basic TA-luc, pCdkn1a-luc, or Bbc3-luc constructs along with the Renilla luciferase plasmid and the TAp63α or RelA or basic vector for 24 h. Immunoblotting for each sample was performed for TAp63α, RelA, and β-actin. Firefly luciferase activity was normalized for the Renilla luciferase activity. Data presented as relative-fold change units (RFU) from the basic TA-luc activity are designated as 1. *, p ≤ 0.001(n = 5) compared with basic TA-luc activity, or pCdkn1a-luc (C) or Bbc3-luc (D) activities alone.