Fig 5.
Wnt target gene activity and alternative activating histone marks in the absence of Dot1l enzyme activity. (A) Quantitative RT-PCR analysis of selected known intestinal Wnt target genes in isolated intestinal crypt epithelium. Average fold differences and standard deviations (±SD) from 3 replicate samples were calculated for each gene in mutant cells compared to controls and normalized to β-actin mRNA. (B) Total levels of known activation- and repression-associated chromatin marks in Dot1lfl/fl (Ctrl) and VillinCreER; Dot1lfl/fl (Mut) mouse intestinal epithelial cells, assessed by immunoblotting of histone extracts. Total H3 served as a loading control. (C) H3K79me2, H4k20me, and H3K36me3 ChIP at representative gene loci in crypt epithelial cells isolated from Dot1l-deficient (Mut) and control (Ctrl) mice. Immunoprecipitated DNA was analyzed by quantitative PCR, and results are represented in relation to the enrichment of a known unmarked fragment.