Senescence associated β galactosidase sa β gal kit

Icaritin was purchased from Sigma-Aldrich (MO, United States). Fetal bovine serum (FBS) was purchased from Gibco (Waltham, MA). 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), crystal violet, a cell-based ROS assay kit, N-acetylcysteine (NAC), and a senescence-associated β-galactosidase (SA-β-Gal) kit were obtained from Beyotime Biotechnology (Beijing, China). Annexin V-FITC/PI and propidium iodide (PI) were purchased from KeyGEN BioTECH (Nanjing, China). The primary antibody against γ-H2AX was obtained from Cell Signaling Technology (MA, United States). Primary antibodies against ki67, GAPDH, PARP1, cyclin D1, CDK2, CDK4, Bax, Bcl-2, P53, P21, p-EGFR, EGFR, p-AKT, AKT, and HIF1α were obtained from Proteintech (Rockville, MD, United States). Antibody against horseradish peroxidase (HRP)-conjugated secondary antibodies (goat-anti-mouse or goat-anti-rabbit) were obtained from ZSGB-BIO (Beijing, China).

The following antibodies were used: anti-SRD5A1 (Proteintech, #66329-1-Ig, 1:1,000 dilution), anti-nuclear factor-κB (NF-κB) p65 (Cell Signaling Technology, #8242, 1:1,000 dilution), anti-Phospho-NF-κB p65 (Cell Signaling Technology, #3033, 1:1,000 dilution), anti-VEGF (Proteintech, #19003-1-AP, 1:1,000 dilution), and anti-β-actin (Proteintech, #66009-1-Ig, 1:5,000 dilution). Secondary antibodies included goat anti-Rabbit IgG(H+L) HRP (Fcmacs, #FMS-Rb01, 1:5,000 dilution) or mouse (Affinity, #S0002, 1:10,000 dilution). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) was purchased from Solarbio (Beijing, China), dutasteride was purchased from APExBIO (Houston, TX, USA), propidium iodide (PI) was purchased from XunBei (Nanjing, China), and Annexin-V-FITC was purchased from Biolegend (San Diego, CA, USA). Pierce TM BCA Protein Assay kit, transfection reagent, and RNase A were purchased from YEASEN (Shanghai, China). Senescence-associated β-galactosidase (SA-β-gal) kit, puromycin, and doxycycline were purchased from Beyotime Institute of Biotechnology (Shanghai, China). TRIeasy™ Total RNA Extraction Reagent, complementary DNA synthesis superMix, and SYBR Green Master Mix were purchased from YEASEN (Shanghai, China).