Ne per r nuclear and cytoplasmic extraction reagents

Cells were lysed in boiling Laemmli sample buffer or processed with NE-PER(R) Nuclear and Cytoplasmic Extraction Reagents (Pierce Biotechnology, Rockford, IL) according to manufacturer’s instruction. Protein samples were electrophoresed on SDS polyacrylamide gels and transferred to nitrocellulose membranes at 100 V for 1 hr. Membranes were probed with the indicated antibodies (Supplementary Table 1), which were detected using HRP-based chemiluminescence (ECL, Pierce Biotechnology, Rockford, IL) [56 (link)]. Densitometric analises were performed with Image J software.

Cells were lysed in boiling Laemmli sample buffer or processed with NE-PER(R) Nuclear and Cytoplasmic Extraction Reagents (Pierce Biotechnology, Rockford, IL, USA) according to manufacturer’s instruction. Extracts were loaded on SDS-polyacrylamide gel, transferred on nitrocellulose membrane, and immunoblotted with antibodies listed in Table 2. β-actin antibody and Histone H3 antibody were used as marker proteins for total and nuclear extracts, respectively. Immunocomplexes were visualized with an enhanced chemiluminescence kit (ECL, Pierce Biotechnology, Rockford, IL) and acquired with ChemiDoc Imaging System (Bio-Rad) applying a fixed exposure time for each antibody. Band intensity was analyzed by ImageJ software.