Cell cycle distribution was evaluated 24 h after cell treatment with the respective IC50 of GSH-NS B or GSH-NS D. The occurrence of the so-called sub-G0/G1 peak, which is a distinct cell population characterized by subdiploid DNA fluorescence and might correlate with the internucleosomal DNA fragmentation typical of apoptosis (Pozarowski and Darzynkiewicz, 2004), was also evaluated. Briefly, 1 × 106 HCT116, 1 × 106 HT-29, 1 × 106 DU145, and 1 × 106 PC-3 cells were incubated with 2 µM of the live cell staining Vybrant Dye Cycle Green (Invitrogen) for 30 min at 37 °C. The samples were run on a flow cytometer with 488 nm excitation to measure Vybrant Dye Cycle Green staining and data analysis was performed by FCS Express software version 4 (BD Bioscience, Milano, Italy).
Fcs express software version 4
Manufactured by BD
Sourced in Italy
FCS Express version 4 is a software application for the analysis and visualization of flow cytometry data. It provides tools for data processing, gating, and statistical analysis of flow cytometry experiments.
Automatically generated - may contain errors
2 protocols using fcs express software version 4
1
Cell cycle analysis of cancer cells
2
Apoptosis Assay for Cell Lines
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HCT-116 and KB cell death was investigated using the Dead Cell Apoptosis Kit with allophycocyanin (APC)-Annexin V and Sytox ® Green (Life Technologies, Milan, Italy) with an Accuri C6 flow cytometer. Cells were incubated for 2 h in FFDMEM medium containing 1.0 nM FA-PEG-GNP suspension, cells were then trypsinized, washed with PBS and normalized to 5.0 × 10 5 cells in 2.5 ml of PBS for US treatment. After US exposure, cells were collected into 3 ml sterile centrifuge tubes for 2 h, washed twice with 1 × Annexin-binding buffer at 1500 r.p.m. for 5 min and stained with APC-Annexin V and Sytox Green for 15 min at 37°C and samples underwent flow cytometric analyses. Cell debris with low forward light scatter and side light scatter were excluded from the analyses and a total of 10,000 events were analyzed. Fluorescence was collected at 660 and 530 nm to discriminate APC-Annexin V and Sytox Green signals, respectively. Apoptotic and late apoptotic/necrotic cells were discriminated from viable cells using the FCS Express software, version 4 (BD, Bioscience, Milano, Italy).
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