Transcriptional regulation of Fgf8 by ΔNp63γ.

<p>(A) 5′-end flanking region up to −3,750 bp from the transcription start site (TSS) of the mouse Fgf8 gene. Six consensus sequences for p63 binding in this region are shown as A1−6. (B) Luciferase activities in B16 melanoma cells co-transfected with a luciferase reporter gene construct containing a fragment (−3,750 bp to +51 bp) of the Fgf8 gene and an expression vector for GFP, ΔNp63γ, or TAp63γ. RLA, relative luciferase activity. Error bars indicate s.d. (<i>n</i> = 3 biological replicates). **<i>P</i> < 0.01 (unpaired two-tailed Student's t test). (C) Luciferase activities in B16 melanoma cells co-transfected with luciferase reporter gene constructs containing the indicated fragments ligated to a minimal promoter (miniP) and an expression vector for GFP or ΔNp63γ. Error bars indicate s.d. (<i>n</i> = 3 biological replicates). *<i>P</i> < 0.05, **<i>P</i> < 0.01 vs. GFP. ^#<i>P</i> < 0.05, ^##<i>P</i> < 0.01 vs. WT with ΔNp63γ (unpaired two-tailed Student's t test). (D) ChIP-qPCR using lysates of mouse ES cells transfected with 3×FLAG-tagged ΔNp63γ. The amplicon of each primer set is indicated as P1–8 in the scheme. Negative control primers were designed in the upstream region of <i>Aldh1a2</i> (Cntl). Error bars indicate s.d. (<i>n</i> = 3 biological replicates). *<i>P</i> < 0.05, **<i>P</i> < 0.01 vs. Cntl (unpaired two-tailed Student's t test).