NextSeq 2000 sequencing; MAT2A promotes reprogramming of the androgen response pathway in association with ERG and EZH2 (ChIP-Seq)
MAT2A promotes reprogramming of the androgen response pathway in association with ERG and EZH2 (ChIP-Seq)
Cell line samples (3x106 cells per IP) were exposed to formaldehyde (37%) to cross-link protein-DNA complexes and processed as previously described (Albino et al., 2012; Longoni et al., 2013). Chromatin extracts were immunoprecipitated with antibodies against AR (cat. 06-680, Millipore), H3K4me2 (cat. 9725S, Cell Signaling) and IgG (Millipore) as control. Quantitative real-time PCR was performed using KAPA SYBR Fast qPCR kit (KAPA Biosystems) and primers spanning the region of interest (Table 3). The amount of immunoprecipitated DNA was calculated in reference to a standard curve and normalized to input DNA or subsequently to IgG (Longoni et al., 2013). For comparison of test and control samples, the amount of DNA was then normalized to the control samples.