Anti oct1

Quantitative real-time RT-PCR amplification was performed on RNA extracted from tissue samples or cell lines using RNeasy mini kit (Qiagen). RNA (50ng) was used for each reaction and the result was normalized by co-amplification of 18S RNA. Reactions were performed on an ABI Prism 7700 Sequence Detection System using Taqman one-step RT-PCR reagents. For ChIP assay, cells were formalin fixed, lysed and sonicated. Anti-AR (Santa Cruz), anti-p300 (Santa Cruz), anti-FOXA1 (Abcam), anti-OCT1 (Santa Cruz), anti-LSD1 (Abcam), anti-H3K4me1 (Abcam), antiH3K4me2 (Upstate), anti-H3K4me3 (Abcam), or rabbit IgG (Santa Cruz) were used to precipitate chromatin fragments from cell extracts. Quantitative real time PCR was used to analyze binding to the ABS1 and ABS2. We used real time quantitative PCR (SYBR green) to amplify the DNA fragment in the antibody precipitated DNA and the un-precipitated input DNA to calculate ΔCT values. The RQ values (RQ=2−ΔCT) are presented and reflect the precipitated DNA as a percentage of the input DNA. Results are represented as mean ± STD for replicate samples. Data are representative of at least three experiments. Significant differences are indicated (*) in the experiments.