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. 2009 May 26;106(23):9280–9285. doi: 10.1073/pnas.0901184106

Fig. 3.

Fig. 3.

WSTF coactivates the transcriptional properties of cardiac transcription factors. (A) Coimmunoprecipitation of WSTF with WINAC and WISH complex components in MEF cells. (B) WSTF and Brg1 cooperatively coactivate the transactivation function of cardiovascular activators on the Gja5(Cx40) promoter (23). Values are mean ± SD for triplicate luciferase assays in MEFs. (C) Knock-down of WINAC components by siRNA lowered endogenous Gja5(Cx40) gene expression estimated by qRT-PCR in cardiogenic-P19 cells. (D) Recruitment of WINAC components on the target gene promoter at E9.5. In vivo ChIP assays were performed as described in Material and Methods. For the Re-IP assay, the immunoprecipitates (IP) and their supernatants (sup.) were sequentially applied for the following ChIP analysis (13). (E) WINAC components were recruited to the Gja5(Cx40) promoter exclusively in E18.5 hearts, but not in livers. ChIP assays were performed with the indicated tissues in both wild-type mice (WSTF+/+) and WSTF homozygous mice (WSTF−/−). The Right image shows a qPCR analysis using the same chromatin samples from E18.5 hearts shown in the Left image using specific primers for qPCR.