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Real-time quantitative reverse transcription PCR analysis of gene expression of DNA methylation related genes.

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posted on 20.02.2013, 00:18 by Justin N. Treas, Tulika Tyagi, Kamaleshwar P. Singh

Total RNA isolated from RWPE-1 cells with chronic exposure to arsenic and/or estrogen was used to perform one-step real-time quantitative reverse transcription PCR as described in materials and methods. Cycle threshold value (Ct value) of each gene was normalized to the Ct value of housekeeping gene GAPDH obtained from the same sample. The gene expression in fold- change was calculated and histogram was plotted using the means of triplicate values. Statistically significant change in gene expression in treated groups as compared to the vehicle treated control is indicated by an *. Similarly, † indicates a significant difference (p<0.05) between As alone and As in combination with E2. ‡ indicates a significant difference (p<0.05) between E2 alone and E2 in combination with As.