Enhanced chemiluminescence ecl solution

Naringin, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), propidium iodide (PI), and Trypan Blue were purchased from Beyotime Biotechnology (Shanghai, China). Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM), Lipofectamine 2000, and TRIzol were obtained from Invitrogen (Carlsbad, CA, USA). RT-PCR kits were obtained from Promega (Beijing, China). SYBR Premix Ex Taq reagents were obtained from TaKaRa (Dalia, China). Anti-Zeb1, anti-cyclin D1, anti-bcl-2, anti-MMP2, and anti-GAPDH antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). HRP-conjugated secondary antibody, BCA protein assay kit, and enhanced chemiluminescence (ECL) solution were purchased from Beyotime Biotechnology. All experiments were completed in the Central Laboratory of our hospital.

The dried fruit of A. villosum was purchased from Yangchun City Spring Gold Agriculture Investment Co., Ltd. (Yangchun, China) and identified by Dr. Zhengming Qian (Dongguan HEC Cordyceps R&D Co., Ltd., Dongguan, China). The voucher specimen (ZMU‐20) was deposited in the research and development center. Assay kits for aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), low‐density lipoprotein cholesterol (LDL‐C), high‐density lipoprotein cholesterol (HDL‐C), glutathione (GSH), glutathione peroxidase (GSH‐Px), total cholesterol (TC), and triglycerides (TG) were obtained from Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). ELISA kits for rat tumor necrosis factor‐α (TNF‐α) and interleukin 6 (IL‐6) were purchased from Nanjing Senberga Biotechnology Co., Ltd. (Jiangsu, China). The bicinchoninic acid (BCA) protein assay kit, radioimmunoprecipitation assay (RIPA) lysis buffer, and enhanced chemiluminescence (ECL) solution were obtained from Beyotime (Shanghai, China). Primary antibody against estrogen receptor 1 (ESR1) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), while anti‐nuclear receptor subfamily 3 group C member 1 (NR3C1) and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). All reagents used in this study were of analytical grade.