Rabbit anti human cyclin d1

Cells were treated with 100 μM SA12 for 48 hours and then lysed with RIPA lysis buffer (Sangon). Untreated cells served as control. The obtained proteins were subjected to 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes (Millipore Corp, Billerica, MA, USA). After blocking with 5% nonfat milk in Tris-buffered saline solution containing 0.05% Tween-20, the membranes were incubated with primary antibodies at 4°C overnight. And then, the membranes were incubated with secondary antibodies conjugated to horseradish peroxidase at room temperature for 2 hours. Signals were visualized by electrochemiluminescence detection, and the density of the blots was analyzed and quantified by Gel-Pro Analyzer 4.0 software. The primary antibodies used for Western blot analysis were rabbit antihuman MECP2, rabbit anti-human cyclin D1, rabbit antihuman CDK4, rabbit antihuman p16 (Abcam, Cambridge, MA, USA), and mouse antihuman β-actin (Cell Signaling, Danvers, MA, USA). The secondary antibodies were horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (IgG) and goat anti-mouse IgG (Zhongshanjinqiao, Beijing, China).

HeLa, SiHa, MDA-MB-231 and L929 cells were provided by the Affiliated Hospital of Qingdao University. C-phycocyanin was purchased from Taizhou Binmei Biotechnology Co., Ltd., Taozhou, China. Carboxymethyl chitosan (CMC) was purchased from Qingdao Honghai Bio-Tech Co. Ltd., Qingdao, China. CD59-specific ligand peptide was synthesized by Chinese Peptide Company, Hangzhou, China. CCK8 was purchased from Beijing Solarbio Science & Technology, Beijing, China. Immunofluorescence staining kit was purchased from Boster Biological Engineering Co., Ltd. Rabbit antihuman cyclinD1, Bcl-2, caspase-3 and cleaved caspase-3 monoclonal antibodies were purchased from Abcam Company. All other chemicals reached analytical grade.