2 aminoethyl diphenylborinate 2 apb

Generic Equivalent to Xylocaine^® (preservative free lidocaine HCl, 1%, 10 mg/mL parenteral solution) was purchased from Hospira Inc. (Lake Forest, IL, USA). The lidocaine was diluted into the culture medium accordingly. GdCl₂, tetracycline, and 2-Aminoethyl diphenylborinate (2-APB) were purchased from Sigma-Aldrich (St. Louis, MO, USA), while 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Fura-2-acetoxymethyl ester (fura-2AM) were purchased from Abcam (Cambridge, UK).

Hyperoside (purity ≥ 98%, Lot. No. JOT-10,193) was ordered from Chengdu Pufei De Biotech Co., Ltd (China). 2-Aminoethyl diphenylborinate (2-APB) and lipopolysaccharides (LPS) were obtained from Sigma-Aldrich (USA). Calf thymus DNA (ctDNA), Lipofectamine 2000, Dulbecco’s Modified Eagle’s Medium (DMEM), and penicillin/streptomycin were products from Thermo Fisher Scientific (USA). Fetal bovine serum (FBS) was ordered from Nobimpex (Germany). 2′,3′-Cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) sodium and 5,6-dimethyl-9-oxo-9 H-xanthene-4-acetic acid (DMXAA) were products from MedChemExpress (USA). Evans blue dye was purchased from Solarbio (China).

Cell culture reagents were obtained from Gibco-Invitrogen (Grand Island, NY, USA). DIDS, staurosporine (STS), histamine, 2-Aminoethyl diphenylborinate (2-APB), flufenamic acid, gramicidin (Gram), Ac-DEVD-AMC (caspase-3 substrate), Ac-DEVD-CHO (caspase-3 inhibitor), Ac-LEHD-AFC (caspase-9 substrate), Ac-IETD-CHO (caspase-8 inhibitor), DMSO, Hoechst 33258 and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ac-IETD-AMC (caspase-8 substrate) was obtained from Enzo Life Sciences (Lausen, Switzerland). Fura2-AM and DiBaC₄ (3) (Bisoxonol) were obtained from Invitrogen (Molecular Probes, Eugene, OR, USA). z-VAD-fmk was purchased from Promega (Madison, WI, USA). Mouse anti-cytochrome c monoclonal antibody (Clone 7H8.2C12) and mouse anti-human PARP (Clone 4C10-5) were purchased from BD-Pharmingen (Chicago, IL, USA). Mouse anti-Caspase-3 (Clone 3G2) was obtained from Cell Signaling (Danvers, MA, USA). Mouse anti-β-tubulin was acquired from ZYMED (Carlsbad, CA, USA). Importantly, DIDS was prepared as a 50-mM stock in dehydrated DMSO, to avoid the hydrolysis of isothiocyanate groups.^{34 (link)} In control studies, 1% dehydrated DMSO neither induced caspase-3 activity nor blocked STS-induced caspase-3 activity (Supplementary Figure S6).

The normal human hepatocyte cell line (L02) was purchased from the Type Culture Collection of the Chinese Academy of Sciences, Shanghai, China. The L02 cells were cultured in complete RPMI-1640 medium (Gibco, Thermo Fisher Scientific, USA) containing 10% fetal bovine serum (FBS, 11011-861, EVERY GREEN) and 1% penicillin-streptomycin (10 kU/mL-10 mg/mL, FG101-01, Transgene, China) and maintained in an incubator with 5% CO₂ at 37°C. The culture medium was renewed every two days, and cells were passaged at 80% confluence using 0.25% trypsin-EDTA. Emodin was purchased from EFEBIO Company (E054323, 98% (HPLC), China). (2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (MitoTEMPO or MT, SML0737, ≥98% (HPLC)) and 2-aminoethyl diphenylborinate (2-APB, #9754, 97% (HPLC)) were from Sigma-Aldrich (China). N-Acetyl-L-cysteine was acquired from Beyotime Biotechnology (NAC, S0077, >99%, China). Tunicamycin (TM) and doxorubicin (DOX) were purchased from Solarbio Life Science (T8480 and D8740, China), and their purities were both more than 98%.

n-Octanol (99% analytical grade, AR), n-octanoic acid (99.0% AR), n-decanol (98%), and n-decanoic acid (99%, AR) were obtained from Macklin Biochemical Co., Ltd., Shanghai, China. Absolute ethanol was provided by Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. 2-Aminoethyl diphenylborinate (2-APB, 97%) was sourced from Sigma Aldrich, Merck, Germany. Reference standards, including caffeic acid and rosmarinic acid (both ≥98% purity), were purchased from Chengdu DesiTe Biological Technology Co. Ltd., Chengdu, China. The salviaflaside standard was prepared at the laboratory of Hunan University of Chinese Medicine. Methanol was purchased from TEDIA (Fairfield, OH, USA). Purified water was prepared using a Mili-Q water purification system (Millipore, Burlington, MA, USA). All other reagents were of analytical quality and acquired commercially.