Rabbit anti ar

To measure ATM protein expression, nuclear lysates were isolated with an NE-PER Nuclear and Cytoplasmic Extraction Reagents kit (Thermo) with protease and phosphatase inhibitor (Thermo Fisher) included. Total protein lysates were extracted by RIPA buffer (Sigma). Protein concentrations were measured using the Bradford reagent (Bio-rad). Equal amounts of protein lysate were loaded onto a 4–15% SDS/PAGE pre-cast TGX Stain-Free Gel (Bio-Rad) and subsequently transferred onto PVDF membrane (Bio-Rad) at 30 V at 4 °C overnight. Immunoblot analysis was performed with the following antibodies: rabbit-anti-ATM (Novus, NB#100–104), mouse-anti-N-MYC (Santa Cruz, #SC-53993), rabbit-anti-NSE (Millipore, # 2716128), rabbit-anti-PSA (Dako, # 20035991), mouse-anti-CgA (Millipore, # MAB5268), rabbit-anti-AR (Millipore, #06–680), mouse-anti-GAPDH (Abcam, #ab9484), rabbit-anti-p84 (Invitrogen, #PA5–27816), rabbit-anti-EZH2 (Cell Signaling Technology, #5246). Following incubation with the appropriate secondary horseradish antibodies (Bio-Rad), blots were developed using the Chemiluminescent Substrate kit (Thermo).