ChIP/quantitative real-time PCR for analyses of the acetylated histones H3 and H4 at the απ-globin promoter in day 5 and day 14 embryonic chicken erythroid cells.
(A) Standard curve generated using varying amounts of chicken genomic DNA. X-axis shows log ng of chicken genomic DNA used as template, and y-axis shows the threshold cycle (CT) value. Based on the standard curve, a linear regression equation was determined. This equation was used to calculate the amount of input DNA from CT values. (B) Bound-unbound ratio for control (chromatin undergoing all the steps in ChIP assay but without addition of an antibody), acetylated histone H3, and acetylated histone H4. Results for acetylated histones H3 and H4 have been normalized for control. Error bars indicate results obtained with 3 independent experiments. For each experiment, real-time PCR was performed in triplicate, and a mean of CT values was used for calculation of the input DNA.
