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. 2009 Mar 20;37(9):3007–3020. doi: 10.1093/nar/gkp179

Figure 7.

Figure 7.

Dazap2 knockdown decreases binding of TCF-4 to the promoters of the Wnt-signalling target genes. STF 293 cells were transfected with Dazap2 siRNA #2 (siDazap2) or control siRNA (siCTRL). After a 2-day expansion the cells were transferred to large culture dishes and stimulated with either Wnt3a ligand or Wnt3a storage buffer only. After 16 h, the cells were harvested and used for mRNA isolation and cDNA preparation. Alternatively, the cells were fixed directly in the dishes and utilized for chromatin immunoprecipitation (ChIP). The left panel shows the results from qRT–PCR analysis. The relative abundance of the indicated mRNAs in Wnt3a-stimulated versus control cells was derived from the average CT values after normalizing to the levels of β-ACTIN cDNA. The right panel shows the ChIP analysis of chromatin isolated from STF 293 cells transfected with control or Dazap2 siRNA #2 duplexes. Only results for Wnt3a-stimulated cells are shown. The diagram represents the relative amounts of the respective DNA element pulled-down by the indicated antibodies. The amount of input non-immunoprecipitated DNA (evaluated separately for each primer set) was arbitrarily set to 100%. Data from one representative experiment from two in total are given.