EtOH blocks RNA polymerase II and MZF1-mediated TGF-β1 promoter activation and RNA polymerase II-mediated MZF1 promoter activation.
A, binding of RNA pol II and MZF1 to the TGF-β1 promoter. ChIP Real Time-PCR was performed using primers that included the TGF-β1 promoter TATA box to determine the role of OPN and EtOH in transcriptional regulation of TGF-β1 expression. B, graphical representation of RNA pol II and MZF1 binding to the TGF-β1 promoter. The percent input was calculated from ChIP Real Time-PCR CT values using input samples as internal controls for the amount of chromatin. C, binding of RNA pol II and MZF1 to the TGF-β1 third exon. ChIP Real Time-PCR was performed using primers that included the TGF-β1 third and final exon to determine the role of OPN and EtOH in transcriptional regulation of TGF-β1 expression. D, graphical representation of RNA pol II and MZF1 binding to the TGF-β1 third exon. The percent input was calculated from ChIP Real Time-PCR CT values using input samples as internal controls for the chromatin amount. E, binding of RNA pol II to the MZF1 promoter. ChIP Real Time-PCR was performed using primers that included the MZF1 TATA box. F, graphical representation of RNA pol II binding to the MZF1 promoter. The percent input was calculated from ChIP Real Time-PCR CT values using input samples as internal controls for the amount of chromatin. Data are presented as mean ± S.E. of three experiments. Gels are representative of three experiments.