Epics xl mcl fcm

Cells in the logarithmic phase were collected and incubated in 6-well plates at a density of 1×10⁵ cells/well. Cells were digested with EDTA-free trypsin (Beyotime Institute of Biotechnology), stained with Annexin V-fluorescein isothiocyanate and propidium iodide (MedChem Express LLC, Monmouth Junction, NJ, USA), and incubated in a dark place for 15 min at room temperature. The apoptosis rates of each group were detected using a flow cytometer (EPICS XL-MCL FCM; Beckman Coulter, Inc., Brea, CA, USA) with an excitation wavelength of 488 nm and an emission wavelength of 530 nm. Data was analyzed using FCS Express version 3.0 (De Novo Software, Glendale, CA, USA).
Cells were treated for 2 h with the lysosomal inhibitors E64d and pepstatin A (10 mg/ml; MedChemExpress USA) or dimethyl sulfoxide (DMSO) following transfection. Monodansylcadaverine (MDC) powder was purchased from Sigma-Aldrich; Merck KGaA and dissolved in DMSO at 0.1 mol/l of the stock concentration. The working concentration was 50 µm/l. The cells were incubated with the MDC dye for 45 min in the dark at 4°C. The cells were subsequently washed with PBS three times. The positive rate of autophagy was determined using a flow cytometer (EPICS XL-MCL FCM; Beckman Coulter, Inc.). Data was analyzed using FCS Express version 3.0 (De Novo Software, Glendale, CA, USA).

The cells were digested by 0.25% trypsin-EDTA (Beyotime Institute of Biotechnology, Haimen, China) and collected by centrifugation at 500 × g at 4°C for 5 min and washed three times with phosphate-buffered saline (PBS). The precipitated cells were resuspended and fixed in 70% absolute alcohol. Subsequently, for the purpose of analyzing cell cycle, the cells were washed with PBS and centrifuged at 500 × g at 4°C for 5 min to collect the precipitate. Propidium iodide (PI) was added for staining. The cell cycle status was determined using an EPICS XL-MCL FCM system (Beckman Coulter, Inc., Brea, CA, USA).
Cells in the logarithmic phase were collected and seeded into 6-well plates (3×10⁵/well). The cells were then digested in EDTA-free trypsin (Shanghai Lanpai Biotechnology Co., Ltd., Shanghai, China), and stained with Annexin V-FITC and PI (Shanghai Lanpai Biotechnology Co., Ltd.). The cells were then incubated in the dark for 15 min at room temperature. The apoptotic rate of the cells in each group was detected using the EPICS XL-MCL FCM (Beckman Coulter, Inc.) with an excitation wavelength of 488 nm and emission wavelength of 530 nm.