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Figure 6 | BMC Molecular Biology

Figure 6

From: Upstream stimulatory factors are involved in the P1 promoter directed transcription of the AbetaH-J-J locus

Figure 6

Chromatin immunoprecipitation assay performed to evaluate in vivo USF interaction with P1 promoter. (A) Representative quantitative real-time PCR profiles for the amplification of the P1 promoter in the ChIP assay. USF1 ChIP, USF2 ChIP and MEF-2A ChIP represents duplicate amplification curves from chromatin immunoprecipitated with USF1, USF2 or MEF-2A antiserum respectively; nonimmune serum represent curves from immunoprecipitations with nonimmune rabbit serum. Input represents curves obtained from HeLa chromatin (1%) before immunoprecipitation. TF, threshold fluorescence. (B) In vivo association of USF1 and USF2 transcription factors with the AβH-J-J P1 promoter. The results, obtained from ChIP assay quantitative real time PCR using USF1 (USF1 ChIP) or USF2 (USF2 ChIP) antiserum, were analyzed following the methodology described in the methods section. The fold increase over negative control PCR in each case compares the values obtained by P1 promoter amplification to the corresponding amplification of a distal genomic region lacking USF1 binding sites (Table 1). All data represent the mean of two separated PCR experiments each performed in triplicate, and obtained from at least two independent immunoprecipitation. The asterisk indicates that the value is significantly different (p < 0,05) from the control value.

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