Ab106693

To obtain total protein extracts from each group, cells were washed with ice-cold PBS and lysed with RIPA lysis buffer (Beyotime, Shanghai, China) on ice. Cells were homogenized and centrifuged. Supernatants were collected and boiled with loading buffer. Cell lysates were subjected to 10% dodecyl sulfate polyacrylamide gels and proteins were transferred onto polyvinylidene difluoride membranes. The membranes were blocked with 5% non-fat milk and incubated with anti-PTEN (ab32199, 1: 1/10,000, Abcam, USA), anti-Bax (ab32503, 1: 2,000, Abcam, USA), anti-Bcl-2 (ab32124, 1: 1,000, Abcam, USA), anti-AKT (ab106693, 1: 1,000, Abcam, USA), anti-PI3K (ab151549, 1: 1,000, Abcam, USA), anti-PI3K p85 (ab191606, 1: 1,000, Abcam, USA), anti-microtubule associated protein 1 light chain 3 (LC3)B-II/I (ab51520, 1: 3,000, Abcam, USA), and anti-GAPDH (ab9485, 1: 2,500, Abcam, USA) antibodies. The membranes were washed and then incubated with horseradish peroxidase–-conjugated secondary antibodies (ab6721, 1: 2,000, Abcam, USA). The resulting light emission was detected with Novex® ECL Chemiluminescent Substrate Reagent Kit (Thermo Fisher Scientific, Massachusetts, USA) and documented with a ChemiDoc™ XRS+ imaging system (Bio-Rad, California, USA).

Roswell Park Memorial Institute 1640 (RPMI 1640) medium (61870-127), fetal bovine serum (26400-036), penicillin-streptomycin (15140-122), and trypsin (25200-056) were purchased from Gibco, USA. The luciferase (L7840) detection kit was purchased from solarbio. The BCA protein concentration determination kit (P0012S), Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit (C1062S), and terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) apoptosis detection kit (C1091) were bought from Shanghai Biyuntian Biotechnology Institute. The Anti Ki67 (ab15580), proliferating cell nuclear antigen (PCNA) (ab18197), Bax (ab53154), Bcl-2 (ab196495), Caspase-3 (ab13847), cleaved caspase-3 (ab2302), Caspase-9 (ab52298), cleaved Caspase-9 (ab2324), phosphatidylinositol 3 kinase (PI3K) (ab191606), p-PI3K (ab182651), protein kinase B (AKT) (ab106693), p-AKT (ab192623), mTOR (ab2732), and β-Actin (ab8227) were purchased from Abcam, USA.