Annexin 5 fitc propidium iodide apoptosis detection kit

RPMI 1640 and pancreatin were from Gibco (USA). Fetal bovine serum (FBS) was from TransGen (China). Trizol^TM reagent was from Life (USA). PrimeScript^TM RT reagent kit with gDNA eraser was from TaKaRa (Japan). FastStart universal SYBR green master and BrdU assay ELISA kit were from Roche (Switzerland). Annexin V-FITC/propidium iodide apoptosis detection kit, Hoechst 33258 and cell cycle detection kit were from KeyGEN (China). Agilent M × 3005P real-time PCR machine was from Agilent (USA). LDN-193189 was from Selleckchem (USA). FACSCalibur flow cytometry was from BD Biosciences (USA). Olympus IX71-A12FL/PH fluorescence microscope and JEOL JEM-1200EX transmission electron microscopy were from Japan. Microplate reader and Nanodrop 2000 were from Thermo Scientific (USA).

Apoptosis and cell cycle rates were assessed using an Annexin V-FITC/propidium iodide apoptosis detection kit and a cell cycle detection kit, respectively (KeyGEN Biotech, Nanjing, China). Quantification of propidium iodide and FITC signals was performed using a fluorescence activated cell sorter FACSAria system (BD Bioscience, NJ, USA).

For cell cycle analysis, briefly, after 12 hours of synchronization by serum starvation, the HepG2 and SNU449 cells transfected with BCL6B expression vector or control empty vector 24 hours before were re-stimulated with 10% fetal bovine serum (FBS) for 24 hours. Cells were fixed in 70% ethanol and stained with 50 μg/ml propidium iodide (BD Pharmingen, San Jose, CA). The cells were then sorted by FACS Calibur (BD Biosciences, Franklin Lakes, NJ) and cell-cycle profiles were analyzed by ModFit version 2.0 (Verity Software House, Topsham, ME) as described in the previous article [7 (link), 39 (link)]. Histograms were analyzed for cell cycle compartments using Early and late apoptosis was detected by Annexin V- FITC/propidium iodide (PI) Apoptosis Detection Kit (KeyGen Biotechnology, China).

H9C2 cardiomyocytes were obtained from the Chinese Academy of Sciences (Beijing, China). Hydrogen peroxide was purchased from Sigma‐Aldrich (MO, USA). Matrigel was purchased from BD Bioscience (NJ, USA). Low‐glucose Dulbecco's Modified Eagle's Medium, foetal bovine serum (FBS), 1% penicillin and streptomycin and 0.05% trypsin‐EDTA were purchased from Gibco (NY, USA). mTeSR1 medium was purchased from StemCell Technologies (BC, Canada). Cell counting kit‐8 (CCK‐8) was bought from BestBio (Shanghai, China). An ABI Prism 5, 59, 6, 69‐tetrachloro‐1, 19, 3, 39‐tetraethylbenzimi‐dazolylcarbocyanine iodide (JC‐1) assay kit, cell cycle kit and Annexin V‐FITC/propidium iodide (PI) apoptosis detection kit were purchased from KeyGEN (Nanjing, China). The fluorescent dye 2′7′‐dichlorofluorescin diacetate assay kit was obtained from Qcbio Science & Technologies (Shanghai, China). An enzyme‐linked immunosorbent assay (ELISA) kit was obtained from Chemicon International (CA, USA). Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were measured using assay kits from Jiancheng Biochemical Inc. (Nanjing, China). SA‐β‐gal staining solution was purchased from the Beyotime Institute of Biotechnology (Nantong, China). The in situ Cell Death Detection kit was purchased from Roche (Basel, Switzerland).

A total of 1 × 10⁶ TDSCs (passage 3) were double‐stained using the fluorescent dye annexin V‐FITC/Propidium Iodide (PI) Apoptosis Detection Kit (Keygen Biotech) according to the manufacturer's instructions. Apoptosis cells (annexin V+/PI‐) were detected using flow cytometry.