Rabbit anti med12

A total of 10–20 μg of whole-cell lysates was resolved by SDS-PAGE and transferred to nitrocellulose membranes (GE Healthcare). Loading of each track was verified with Ponceau S (Sigma-Aldrich) staining. Antibodies used were as follows: mouse anti-AR (Santa Cruz, #7305), rabbit anti-AR (Santa Cruz, #816), rabbit anti-KLF4 (Abcam, #215036), mouse anti-tubulin (Cell Signaling Technology, #3873), mouse anti-actin (Cell Signaling Technology, #3700), anti-GFP (Abcam, #290), rabbit anti-MED1 (Bethyl, #A300–793A), rabbit anti-MED12 (Bethyl, #A300–774A), rabbit anti-KMT2A (Bethyl, #A300–087A), mouse anti-DMAP1 (Santa Cruz, #373949), rabbit anti-POLH (Bethyl, #A301–231A), rabbit anti-NIPBL (Bethyl, #A301–779A), or mouse H3 (Cell Signaling Technology, #3638). Secondary antibodies were the following: anti-FLAG M2 (Sigma-Aldrich), horse anti-mouse HRP-linked IgG (Cell Signaling Technology, #7076), goat anti-rabbit HRP-linked IgG (Cell Signaling Technology, #7074), or streptavidin-HRP (Life Technologies #434323). Signal was revealed using BioRad Clarity Western ECL substrate and detected either on Hyblot CL autoradiography films (Denville) or with an Amersham Imager 600RGB (GE Healthcare). Signal quantification was performed using Image J software gel analysis tools (NIH).