Dutasteride

Caspase-3 (#9662, 1:1000), Cleaved Caspase-3 (#9661, 1:1000), PARP (#9542, 1:1000), Bcl-xl (#2762, 1:1000), Bcl-2 (#2876, 1:1000), Bad (#9292, 1:1000), Bax (#2774, 1:1000), ATG-5 (#12994, 1:1000), ATG-7 (#8558, 1:1000), LC3A/B (#12741, 1:1000), Akt (#9272, 1:1000), p-Akt (Thr308) (#9275, 1:1000), p-mTOR (#2971, 1:1000), p-p70S6K (Thr389) (#9234, 1:1000) and β-actin (#4970, 1:1000) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). SRD5A1 (#66329-1-Ig, 1:1000) and p70S6K (#14485-1-AP, 1:2000) were purchased from Proteintech (Manchester, UK). The p-PI3K (Tyr607) (#AF3241, 1:1000) and mTOR (#AF6308, 1:1000) antibody were purchased from Affinity (Cambridge, UK). SQSTM1/p62 (#P0067, 1:1000) antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA) and goat Anti-rabbit IgG/Alexa Fluor 647 antibody (#bs-0295G-AF647, 1:500) was purchased from Bioss (Woburn, MA, USA). LY2940002 (#9901) were obtained from Cell Signaling Technology. Dutasteride (#A1659), hydroxychloroquine (HCQ) (#B4874) and 3-methyladenine (3-MA) (#A8353) were purchased from APExBIO (Houston, USA). Doxorubicin (#D1515) was purchased from Sigma-Aldrich.

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).