Plasmid Constructs and Antibody Details

The plasmid encoding His-fused full length human PARP1 was a gift from John Pascal and described elsewhere [52 (link), 53 (link)]. The plasmid encoding the HA-fused full length human EZH2 plasmid (pCMVHA hEZH2) was a gift from Kristian Helin (Addgene plasmid # 24230) [54 (link)]. The plasmid encoding GST-fused full length human EZH2 (Pgex-EZH2) was a gift from Mien-Chie Hung (Addgene plasmid # 28060) [55 (link)]. All plasmids were verified by sequencing. Rabbit polyclonal anti-His (WB 1/200, Ip: 2.5 μgr/sample), rabbit anti-Mouse IgG HRP conjugated (WB: 1/5000; IP 2.5 μgr/sample), and mouse anti-Rabbit IgG HRP conjugated (WB: 1/5000, IP: 2.5 μgr/sample) were from Santa Cruz Biotechnology. Monoclonal mouse anti-HA (WB 1/1000) was from Origene. Monoclonal mouse anti-HA (IP 2.5 μgr/sample) and rabbit polyclonal anti-Histone H3 (WB 1/2000) were from Abcam. Rabbit polyclonal anti-PARP1 (WB 1/10000) and rabbit polyclonal anti-H3K27me3 (WB 1:2000) were from Active Motif. Rabbit polyclonal anti-PAR antibody (WB 1/1000) was from Trevigen. Mouse monoclonal anti-EZH2 antibody (WB 1/2000; IP 2.5 μgr/sample) was from BD Bioscience. Rabbit polyclonal anti-Actin antibody (WB 1/100) was from Sigma Aldrich. Anti-mono-ADP-ribose binding reagent and Anti-mono- and poly-ADP-ribose binding reagent (IP 5 μgr/sample) were from Millipore.