Anti lamin b1 sc 20682

Whole cell lysates were obtained using the M-Per Mammalian Protein Extraction Reagent (Pierce Biotechnology, Woburn, MA). Nuclear protein extracts were prepared using a Nuclear Extraction Kit (Chemicon International, Temecula, CA) according to the manufacturer's instructions. Total proteins (40 μg) and nuclear proteins (10 μg) were separated on 10% SDS-PAGE and transferred to nitrocellulose membranes. Antigen-antibody complexes were detected using the enhanced chemiluminescence (ECL) blotting analysis system (Amersham Pharmacia Biotech, Buckinghamshire, UK). The following antibodies were used for analysis: anti-EZH2 (5246, Cell Signaling, Danvers, MA), anti-PARP (9542, Cell Signaling, Danvers, MA), rabbit polyclonal anti-E-cadherin (A01589, GenScript, Edison, NJ), mouse monoclonal anti-N-cadherin (BD, Transduction, San Jose, CA), rabbit polyclonal anti-Vimentin (A01189, GenScript, Edison, NJ). Anti-MCL-1 (sc-819), anti-FOS (sc-52), anti-p21 (sc-397), anti-Bax (sc-493), anti-β-catenin (sc-1496), and anti-GAPDH (sc-47724) and anti-lamin B1 (sc-20682) were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA). GAPDH (whole cell lysate) and lamin B (nuclear protein) were blotted to show equal protein loading, respectively.

All cell culture media, supplements and modifying enzymes were purchased from Invitrogen (Karlsruhe, Germany) if not otherwise indicated. Doxorubicin was from Biotrend Chemicals (Cologne, Germany), paclitaxel, and propidium iodide were obtained from Sigma-Aldrich (Taufkirchen, Germany). Gö6983, KU55933, SB203580, U0126 and Chk2 inhibitor II were from Merck (Darmstadt, Germany). The caspase inhibitors Z-VDVAD-FMK and Z-VAD-FMK were derived from R & D Systems (Wiesbaden, Germany). The following antibodies were used in this study: anti-HuR (sc-5261) and anti-Lamin B1 (sc-20682) from Santa Cruz (Heidelberg, Germany), anti-caspase-2 (#611022, BD Biosciences, Heidelberg, Germany), anti-caspase-3 (#9662), anti-caspase-7 (#9494) anti-PARP (#9542) and anti-Bid (#2002) antibodies were from Cell Signaling (Frankfurt, Germany), and anti-β-actin (#A2228) from Sigma-Aldrich. The goat anti-rabbit (sc-2054) and goat anti-mouse (sc-20559) HRP-linked antibodies were from Santa Cruz and Alexa Fluor 488 goat anti-mouse was from Life Technologies (Darmstadt, Germany). Radionucleotides were from Perkin Elmer (Rodgau, Germany) and the ECL system and Hyperfilms were from GE Healthcare (Freiburg, Germany).