Flow cytometry (FACS)-based assays were performed on A549 GLUL KO cells treated with docetaxel to analyze apoptosis. For apoptosis assay, PE-conjugated annexin V and 7-AAD staining was performed according to the manufacturer’s protocol (BD Biosciences, San Jose, CA, USA). Data were acquired using BD LSR II flow cytometer, and the analysis was performed with FCS Express 6Plus software (De Novo Software, Pasadena, CA, USA).
Fcs express 6 plus software
Manufactured by De Novo Software
Sourced in United States
FCS Express 6 Plus Software is a comprehensive data analysis platform designed for flow cytometry data. It provides advanced tools for data visualization, statistical analysis, and reporting. The software supports a wide range of flow cytometry instruments and file formats, enabling users to efficiently analyze and interpret their experimental data.
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Lab products found in correlation
Celltrace cfse, by Thermo Fisher Scientific (2 mentions)
Cd8 apc cy7, by BD (2 mentions)
Cd3 pe, by BD (2 mentions)
Cd4 apc, by BD (2 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Facscanto, by BD (1 mentions)
Accuri c6 system, by BD (1 mentions)
Celltrace violet, by Thermo Fisher Scientific (1 mentions)
Facscanto flow cytometer, by BD (1 mentions)
Facscalibur flow cytometry system, by BD (1 mentions)
Fluorescein isothiocyanate fitc labeled mouse anti human cd90, by STEMCELL (1 mentions)
6 protocols using fcs express 6 plus software
1
Apoptosis Analysis in A549 GLUL KO Cells
2
Apoptosis Analysis by Flow Cytometry
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Apoptotic cell population was determined by evaluating the DNA content of hypodiploid cells by the PI flow cytometric assay66 (link),68 (link). Cells were seeded, and (or) transfected with individual siRNAs for 48 h. Cells were treated with TM or TG for 48 or 72 h. Cells were fixed with cold 70% ethanol and treated with PI/RNase staining solution as described in the manufacturer’s instructions. Cells were analyzed with a FACSCalibur flow cytometer (BD Biosciences), and cellular DNA content and forward scatter were analyzed with FCS Express 6 Plus software (De Novo Software) or WinMDI2.9 software. To determine PARP1 cleavage, cells were prepared as above, and then immunoblot analyses were performed with anti-PARP1 antibody as recommended by the supplier.
3
T-cell Proliferation Assay with Lentiviral Transduction
4
Apoptosis and Cell Cycle Analysis
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For apoptosis and cell cycle analyses, cells were seeded and allowed to settle overnight, followed by treatment with the indicated substances. After incubation, cells were trypsinized, washed with PBS and stained with propidium iodide. Fluorescence-activated cell sorting was performed using BD Accuri C6 system. The percentage of cells in sub-G1, G0/G1, S or G2/M phase were assessed by FCS Express 6 plus software (De novo software, Pasadena, CA, USA).
5
T-Cell Proliferation Assay Protocol
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For primary T-cell proliferation assays, anti-CD3/CD28 beads were removed five days after lentiviral transduction, and activated T cells (A-T) were left resting for 24 hours at a concentration of 0.8 × 106 cells/mL. Then, transduced or nontransduced A-T were stained with 2.5 μmol/L Cell Trace Violet (catalog no. C34557, Life Technologies) and cocultured with freshly isolated T cells (nonactivated T cells, NA-T) from the same donor, previously stained with 2.5 μmol/L Cell Trace CFSE (catalog no. C34554, Life Technologies), and NALM6 or HeLa target cells at the indicated ratios. After 5 days, samples were stained with CD3-PE, CD4-APC, and CD8-APC-Cy7 (all from BD Biosciences; see Supplementary Table S1) and acquired in a FACSCanto flow cytometer. T-cell proliferation was analyzed using FCS Express 6 Plus Software (De Novo Software).
6
Mesenchymal Stem Cell Characterization
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The cells obtained were checked for their mesenchymal profile by FACSCalibur flow cytometry system (Becton Dickinson, CA, USA). 2.5×10 5 cells were distributed to 5 ml round-bottom polystyrene tubes, washed with Dulbecco PBS and were then stained for 45 min with the following antibodies: fluorescein isothiocyanate-(FITC)-labeled mouse anti-human CD90 (StemCell Technologies, Milan, Italy), CD105, CD14, CD19 and CD34 (Immunotools, Germany), Rphycoerythrin -(PE)-labeled mouse anti-human CD45 (Diaclone, France), CD73 (Becton Dickinson, CA, USA), and HLA-DR (Diaclone, France). The control for FITC-or PE-coupled antibodies was isotypic mouse IgG1. The data were analysed using FCS Express 6 Plus Software (De Novo Software, CA, USA).
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