Parp 1 clone c2 10

Total cell lysates were prepared in RIPA buffer (50 mM Tris-HCl at pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS and 1 mM EDTA) added with sodium orthovanadate 3 mM, NaF 100 mM supplemented with protease inhibitors (Sigma-Aldrich). Lysates were kept on ice for 25 min and then centrifuged at 16000 g for 30 min at 4°C. Supernatants were collected and quantified by Bradford protein assay reagent (Biorad). 25 μg of proteins were loaded and separated by SDS-PAGE and electroblotted onto nitrocellulose membrane (Hybond^TM ECL^TM GE Healthcare). Immunoblots probed with the specific antibodies were developed using ECL or ECL Plus chemiluminescence reaction. The following antibodies were used: Snail (L70G2) (Cell Signaling), PARP-1 (clone C2–10 able to detect the 89kDa cleaved fragment of PARP-1; Enzo Life Sciences), PARP-1 (clone F1–23; Enzo Life Sciences), PAR (clone 10HA; Trevigen), Actin (Sigma-Aldrich), Akt (Cell Signaling), Phospho-Akt (Ser473) (Cell Signaling), PTEN (Cell Signaling), Phospho-Histone H2AX (Ser139) (Millipore).

The following monoclonal antibodies were used: PARP-1 (clone C2-10; Enzo Life Sciences), PAR (clone 10HA; Trevigen), TET1 (Genetex), Myc-tag (9E10 clone, hybridoma-conditioned medium), GST (Thermo), αTUB (Sigma-Aldrich), FLAG (Thermo Scientific Pierce Antibodies). The following polyclonal antibodies were used: PARP-1 (Enzo Life Sciences), PAR (10H, kind gift of A. Burkle), 5hmC (Active motif), Lamin B1 (Abcam).

The following monoclonal antibodies were used: PARP-1 (clone C2-10; Enzo Life Sciences), PARP-2 (Enzo Life Sciences), PAR (clone 10HA; Trevigen), TET1 (Genetex), Myc (9E10 clone, hybridoma-conditioned medium), αTUB (Sigma-Aldrich). The following polyclonal antibodies were used: LMNB1 (Abcam), PARP-1 (Enzo Life Sciences), TET1 (Millipore), PAR (Trevigen), RNA POL II (Santa Cruz Biotech.), H3K4me1 (Millipore), H3K4me2 (Millipore), H3K4me3 (Millipore).