Anti β actin antibody

Cultured cells were harvested and lysed with NP-40 lysis buffer (Beyotime, Haimen, China) for total protein extraction, whereas xenograft tumor tissues were physically homogenized and lysed with RIPA lysis buffer containing 1% v/v PMSF (Beyotime). Proteins were separated by SDS-PAGE, and transferred onto a PVDF membrane (Millipore, Bedford, MA, USA). The membrane was blocked with 5% milk in TBST buffer (TBS + 0.05% v/v Tween-20), and probed for protein of interest with a specific primary antibody overnight at 4°C, followed by incubation with a secondary antibody for 1 h at room temperature (RT). Anti-YB-1 antibody was purchased from Cell Signaling (Boston, MA, USA), anti-cyclin D, anti-cyclin A, anti-cleaved caspase-3, anti-cleaved PARP, anti-Bcl-2 and anti-Bax antibodies were from Wanleibio (Shenyang, China); HRP-conjugated goat anti-rabbit IgG secondary antibody was purchased from Beyotime. Immune complexes were visualized using the ECL system (Qihai Biotech, Shanghai, China). To verify equal protein loading and transfer, membranes were stripped with stripping buffer (Beyotime) and re-probed with anti-β-actin antibody (Wanleibio).

HSYA was purchased from Dalian Meilun Biotechnology Co., Ltd. (Liaoning, China; purity > 98%, powder). HSYA powder was dissolved in sodium chloride as a working solution. D-gal was provided by Sigma-Aldrich (St. Louis, MO, USA). Trizol reagent, Prime Script Master Mix, and SYBR-qRT-PCR Kits were obtained from TaKaRa Biotechnology Co., Ltd. (Dalian, China). BCA Protein Assay Kits were ordered from Bytec Biotech Co., Ltd. (Beijing, China). Anti-HAP90AA1 antibody, anti-ATP2A1 antibody, anti-NOS1 antibody, anti-PKG and anti-β-actin antibody were purchased from Wanlei Biological Technology Co., Ltd. (Shenyang, China). HRP-conjugated goat anti-rabbit IgG/HRP was obtained from Bioss Biotechnology Co., Ltd. (Beijing, China).

Pioglitazone, GW9662, and Aβ_1–42 were purchased from Sigma, USA. Rabbit anti-rat polyclonal antibodies against IDE, BACE1, and APP were purchased from Abcam, UK. Rabbit anti-rat polyclonal antibodies against CDK5, PPARγ and Aβ_1–42 were purchased from Wanleibio, Shenyang, China. Rabbit anti-rat polyclonal antibody against p-PPARγ-Ser273 was purchased from Bioss, Beijing, China. TUNEL assay kit, anti-β-actin antibody, and ECL reagent were purchased from Wanleibio, Shenyang, China. TRIzol, Super M-MLV reverse transcriptase, RNase inhibitor, and 2× Power Taq PCR MasterMix amplification reagent kit were purchased from BioTeke, Beijing, China. SYBR GREEN Master Mix was purchased from Solarbio, Beijing, China. Protein A sepharose beads were purchased from Santa Cruz, CA, USA.