Epics xl mcl facscan

HUVECs were collected and resuspended in fresh culture medium containing 10% FBS. Cells were collected and gently resuspended in 70% ice-cold ethanol for fixing at -20°C overnight. Subsequently, the cells were collected, washed and resuspended in fresh culture medium containing 10% FBS. The cells were then collected and resuspended in 200 µl Cell Cycle Solution [containing propidium iodide (PI), RNase A, and Triton X-100; Invitrogen; Thermo Fisher Scientific, Inc.] at a density of 2×10⁵ cells. Cells were then incubated at room temperature for 30 min without light and finally analyzed using an EPICS XL-MCL FACScan flow cytometer (Beckman Coulter, Inc.).

GC cells (1×10⁶) were washed twice with ice-cold PBS, treated with trypsin and fixed in 70% ethanol at 4°C for 30 min. The cell pellet was then incubated in a solution containing 50 ng/ml propidium iodide, 0.2 mg/ml RNase and 0.1% Triton X-100 at room temperature for 30 min. Subsequently, the cells were analyzed by flow cytometry using an EPICS XL-MCL FACScan device (Beckman Coulter, Inc., Brea, CA, USA). The data were analyzed with MultiCycle software, version 306 for Windows (Phoenix Flow Systems, San Diego, CA, USA).
SGC7901, SNU-1, SNU-5 parental or resistant cells were treated with 20 nM Dox, 40 nM PD and 20 nM LEE011 alone or in combination at 37.8°C for 24 h. Cells were treated with 0.1% DMSO as the control. The apoptosis of GC cells was evaluated by Hoechst 33258 staining. The cells were stained with 10 µg/ml Hoechst 33258 (Invitrogen; Thermo Fisher Scientific, Inc.), in 0.5% Nonidet P-40 and 3.7% formaldehyde at room temperature for 30 min. Subsequently, a microscope was used to observe the cell apoptosis, which was characterized by micronucleation and condensed chromatin in the cells (18 (link)).