Annexin 5 apoptosis kit

Isoliensinine (98% by high performance liquid chromatography; Beijing Solarbio Science & Technology Co., Ltd.) was dissolved in DMSO to 20 mg/ml. 0.1% DMSO was used as the vehicle control for isoliensinine.
AKT inhibitor AKTi-1/2 (10 mM; cat. no. SF2784) was purchased from Beyotime Institute of Biotechnology. The Cell Counting Kit-8 (CCK-8) was purchased from Shanghai Yeasen Biotechnology Co., Ltd. The Annexin V apoptosis kit (cat. no. LHK601-020) was purchased from Beijing Jiamei Nuno Biotechnology Co., Ltd.
Primary rabbit anti-human AKT (cat. no. 4691), phosphorylated (p-)AKT (S473) (cat. no. 4060), p21 (cat. no. 2947), CDK2 (cat. no. 2546), Cyclin E1 (cat. no. 20808), GSK3α (cat. no. 4337), Mcl-1 (cat. no. 94296), cleaved caspase-9 (cat. no. 9508), and GAPDH (cat. no. 5174) antibodies and secondary antibodies were purchased from Cell Signaling Technology, Inc. Secondary antibodies included the HRP-conjugated anti-rabbit antibody (cat. no. 7074) and HRP-conjugated anti-mouse antibody (cat. no. 7076). All primary antibodies were diluted at a ratio of 1:1,000, whilst all secondary antibodies were diluted at a ratio of 1:2,000 for use in the study.

Zinc acetate, 2-methylimidazole (2-MI) and hemin were purchased from Aladdin Biochemical Technology Co. Ltd. (Shanghai, China). Gox and 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) were obtained from Sigma-Aldrich (Shanghai, China). 4′,6-diamidino-2-phenylindole (DAPI) and GSH detection kit were purchased from Solarbio (Beijing, China). Cell membrane fluorescence probe (Dio), an ATP detection kit, BCA protein assay kit and live, dead viability assay kit and Protease inhibitor cocktail (100 × ) were purchased from Beyotime Biotechnology (Shanghai, China). Annexin V apoptosis kit was obtained from Yeason (Shanghai, China). Antibodies against CD45, CD3, CD8, CD4, CD11c, CD80, CD86, IFN-γ for flow cytometry were obtained from BioLegend (San Diego, California, USA). ELISA kits for analysis of cytokines were purchased from Thermo Fisher (Waltham, USA). Anti-mouse-PD-L1 antibody (BE0101) were purchased from Bio X Cell (Lebanon, USA). All other chemical reagents were analytically pure and used directly as received.