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- _5 value "Both RFX1 and RFX3 Down-regulate the MAP1A Promoter in Non-neuronal Cells-- Because EMSA experiments suggested that RFX1 and 3 are the major proteins binding to the RFX sites in the MAP1A gene, we examined their functional effects on the promoter activity in reporter assay. We made RFX1 and RFX3 expression constructs driven by the CMV promoter. The expression of proteins in expected sizes and appropriate nuclear localizations of EGFP-tagged proteins were confirmed by Western blotting and confocal microscopy, respectively (data not shown). We then examined their effects on the hMP1-8 reporter activity in non-neuronal and neuronal cells (Fig. 6A). Overexpression of RFX1 resulted in down-regulation of the reporter compared with control in all non-neuronal cells examined (Fig. 6B, pFC R1). Reporter activity was also down-regulated when RFX 3 was overexpressed in HT29 and HEK293 cells (Fig. 6B, pFC R3). However, overexpression of RFX3 did not result in down-regulation, but slight up-regulation of the reporter in HeLa cells." provenance.