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Cotransfection of CV1 cells with the TKCAT reporter construct and the expression constructs for either MEF2A or MASH1 produced no significant change in reporter gene activity (Fig. 2B). However, coexpression of both MEF2A and MASH1 together resulted in a significant increase in CAT reporter activity. That this induction was due to interaction of the respective transcription factors with their DNA binding sites was supported by the demonstration that point mutations in either the E box or the MEF2 binding site, which in DNA mobility shift assays disrupted MASH1 or MEF2A binding, respectively (data not shown), rendered the reporter gene unresponsive to both MASH1 and MEF2A individually or in combination (Fig. 2B).
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