Quantitation of pulldown was performed by real time PCR using 2× DyNAmo syprogreen qPCR kit (New England Biolabs) according to manufacturers procedures. Briefly, reactions were composed of 5 µl 1.2 µM forward and reverse primers, 5 µl diluted cDNA template and 10 µl 2× qPCR mix (see Table S3 for primer sequences). Reactions were heated for 10 min at 94°C, then cycled for 40 cycles at 20 sec at 94°C, 20 sec at 60°C, 30 sec at 72°C. After each cycle the sample was heated to 78°C for 10 sec prior to reading sample fluorescence. Pulldowns were quantified as a percentage of input using a dilution series as a standard curve. Histone modification pulldowns are expressed as enrichment relative to histone H3 occupancy under +activin-A conditions. SNF2H/L pulldowns were first normalized to signal at Gapdh and are expressed as the enrichment during +activin-A relative to −activin-A conditions.
