Comparison of FIZ1 recruitment and activated Pol-II association, at the Rhodopsin gene in P-3 and P-25 retina. ChIP was performed using antibodies specific for FIZ1 and the actively transcribing form of RNA Polymerase-II (Pol-II-S2). Protein binding is proportional to the amount of target DNA captured by ChIP, and is shown as copies of target DNA detected per 1000 cell equivalents of input DNA. Results were normalized for chromatin input and PCR efficiency. Error bars indicate standard deviation (n = 3). A) FIZ1: Q-PCR of two targets in the Rho proximal promoter (-711, -12), as illustrated in the diagram of the Rho gene. An untranslated region (Untr6), on chromosome-6, provided a reference for binding (negative control), and was not different comparing P-3 to P-25 neural retina. FIZ1 recruitment to the Rho promoter was significantly increased in P-25 neural retina compared to P-3 retina. B) Pol-II-S2: Q-PCR of a target inside intron-2 of the Rho gene (+3051), and the untranslated region (Untr6). Results confirm an increased amount of actively transcribing Pol-II in the transcriptional region of Rho in P-25 retina compared to P-3 retina.
Mali et al. BMC Molecular Biology 2008 9:87 doi:10.1186/1471-2199-9-87