Fasc calibur mt flow cytometer

Flow cytometry was performed to detect the ratio of p75NTR⁺ cells in ESCC cells. Briefly, Cells were trypsinized and collected by centrifugation. Then cells were washed and suspended in buffer and incubated with p75NTR-PE antibody (BD Bioscience) at 4°C for 2h. The ratio of p75NTR⁺ cells in each samples were analyzed by FASC Calibur MT flow cytometer (BD Bioscience).
Cell cycle was detected by flow cytometry using a cell cycle analysis kit (BD Bioscience). In brief, at least 1*10⁶ cells were harvested and washed, then cells were fixed in ice-cold 70% ethanol for at least 2h at 4°Cwashed the cells again and stained with a solution containing 50μg/ml PI and 50 RNase at room temperature for 30 min. Cell cycle was analyzed with a FACS Calibur MT flow cytometer (BD Bioscience).
Cell apoptosis was detected by flow cytometry using an Annexin V-APC apoptosis detection kit (BD Bioscience). In brief, cells were harvested and washed, then cells were suspended in binding buffer and incubated with Annexin V-APC and propidium iodide (PI) at 4°C, the cells were double stained with Annexin V-APC and PI according to the manufacturer's instructions. Early apoptosis and the late apoptosis were determined by Annexin V+/PI- staining and Annexin V+/PI+ staining, respectively. The percentage of apoptosis cells in each sample was examined using the FASC Calibur MT flow cytometer (BD Bioscience).

To detect cell apoptosis, 16HBE cells were seeded in six-well plates (Corning, NY, USA) and divided into the following four treatment groups: PNS-R1 + Dex group, PNS-R1 group, Dex group and control group. Cells were treated with Dex (5 μM) and PNS-R1 (2 μM) for 24 h. Apoptosis was measured by flow cytometry using the Annexin V-PI apoptosis-detection kit (KeyGen BioTech, China) according to the manufacturer’s instructions. To detect the cell mitochondrial membrane potential (Δψm), we also used the JC-1 apoptosis-detection kit (KeyGen BioTech, China). The percentage of apoptotic cells was calculated using the FASC Calibur MT flow cytometer (BD Bioscience, NJ, USA).

Apoptosis was determined by flow cytometry analysis using the Annexin V-FITC/PI Apoptosis Kit (Becton Dickinson, Franklin Lakes, NJ, United States). Cells were seeded in 6-well plates (5 × 10⁵ cells/well), digested with trypsin (Gibco trypsin-EDTA; Thermo Fisher Scientific), washed with phosphate-buffered saline (PBS) three times, suspended in 500 μl of binding buffer, and then incubated with 5 μl of fluorescein isothiocyanate (FITC)-conjugated Annexin V and 3 μl of propidium iodide (PI) for 15 min at room temperature in the dark. After incubation, the samples were tested using a flow cytometer (Moflo XDP, United States). Early apoptotic chondrocytes were defined as annexin V-positive and PI-negative cells, and late apoptotic chondrocytes were defined as annexin V-positive and PI-positive cells. The proportion of apoptotic cells was calculated using the FASC Calibur MT flow cytometer (BD Bioscience, NJ, United States).