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. Author manuscript; available in PMC: 2010 Jul 1.

Published in final edited form as: Nat Genet. 2009 Dec 20;42(1):89–93. doi: 10.1038/ng.494

Wt1 is an activator of Snai1 and a repressor of Cdh1 in epicardial cells. (a, e) Schematic representation of the putative conserved Wt1 binding sites (, □) in the Snai1 and Cdh1 loci, (functional binding site), □ (putative but non functional binding site) and ■ (exons). (b) Luciferase activity of reporter construct carrying mouse Snai1 promoter in epicardial cells in the presence of indicated amounts of −KTS Wt1 expression vector. (c) Luciferase activity of wild-type (Control) or mutated Snai1 promoters in the presence of -KTS Wt1 isoform. (d, f) Cell extracts from epicardial cells were chromatin immunoprecipitated (ChIP), using antibodies against Wt1, anti-acetyl-histone H3 (AcH3), anti-H3 tri methyl K4 (K4Me) and anti- H3 tri methyl K27 (K27Me) or an irrelevant antibody (IgG). The input was used as a positive control for PCR of the Snai1 (d) and Cdh1 (f) promoters, intronic regions and 3′UTRs. (g, h) Luciferase activity of constructs carrying DNA fragments identified by the Cdh1 ChIP assay in epicardial cells, together with different concentrations of −KTS Wt1 isoform. (i) Luciferase activity of Control or mutated Cdh1 constructs in the presence of -KTS Wt1 isoform (100ng). (j) ChIP assays of Snail and Wt1 at the endogenous Cdh1 promoter in epicardial cells. The graphs represent the mean values ±s.e.m from three independent experiments.

Inline graphic — Wt1 is an activator of Snai1 and a repressor of Cdh1 in epicardial cells. (a, e) Schematic representation of the putative conserved Wt1 binding sites (, □) in the Snai1 and Cdh1 loci, (functional binding site), □ (putative but non functional binding site) and ■ (exons). (b) Luciferase activity of reporter construct carrying mouse Snai1 promoter in epicardial cells in the presence of indicated amounts of −KTS Wt1 expression vector. (c) Luciferase activity of wild-type (Control) or mutated Snai1 promoters in the presence of -KTS Wt1 isoform. (d, f) Cell extracts from epicardial cells were chromatin immunoprecipitated (ChIP), using antibodies against Wt1, anti-acetyl-histone H3 (AcH3), anti-H3 tri methyl K4 (K4Me) and anti- H3 tri methyl K27 (K27Me) or an irrelevant antibody (IgG). The input was used as a positive control for PCR of the Snai1 (d) and Cdh1 (f) promoters, intronic regions and 3′UTRs. (g, h) Luciferase activity of constructs carrying DNA fragments identified by the Cdh1 ChIP assay in epicardial cells, together with different concentrations of −KTS Wt1 isoform. (i) Luciferase activity of Control or mutated Cdh1 constructs in the presence of -KTS Wt1 isoform (100ng). (j) ChIP assays of Snail and Wt1 at the endogenous Cdh1 promoter in epicardial cells. The graphs represent the mean values ±s.e.m from three independent experiments.