Notum is a positive target of β-catenin (original) (raw)

2016

Abstract

(A) Total RNA was extracted from 96 samples (29 from HCC specimens with WT CTNNB1, 38 from HCC specimens with mutant CTNNB1 and 29 from adjacent non tumor tissue that included 19 CTNNB1-mutated HCC and 10 non-mutated HCC). Notum mRNA was assayed by quantitative RT-PCR. (B) NOTUM in situ hybridization with mutantβ-catenin HCC specimens showed significantly stronger NOTUM labeling in tumor tissue than normal tissue. Seven HCC biopsies were analyzed and a representative result is shown. (C) Tamoxifen-inducible model of acute β-catenin activation in the liver. (D) Eight week-old mice were intraperitoneally injected with 0.3 mg tamoxifen and total RNA was extracted from WT liver (black rows) and from APCΔex14/Δex14 liver (grey rows) at the indicated times and Notum mRNA was assayed by quantitative RT-PCR. Results are normalized to those for 18S transcripts and the mean ± S.D. of triplicate samples is presented. (E) Genomic environment of the Notum gene (from mRNA-Seq and ChIP-Seq data described in [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150997#pone.0150997.ref005" target="_blank">5</a>]. Upper blue: mRNA-seq signals. ChIP-Seq signals are shown below with the presence within peaks of WREs depicted first in red; then in black, ChIP-Seq in Apc-ko conditions with an antibody against Tcf4; in red, ChIP-Seq in Apc-ko conditions with an antibody against β-catenin; in green, ChIP-Seq in β-catenin-ko conditions with an antibody against Tcf4. Below in yellow, ENCODE data for DNase 1 HS performed with 8-week-old and in E14.5 embryonic mouse livers (DNaseI Hypersensitivity by Digital DNaseI from ENCODE/University of Washington), of H3K27Ac marks in 8-week-old and in E14.5 embryonic livers (Histone Mods by ChIP-seq from ENCODE/LICR).</p

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