Cells were seeded and subsequently treated with selected inhibitors over a period of 24 h. We then fixed the cells using cold 70% ethanol and treated them with ribonuclease. Cell cycle-dependent uptake of PI was observed using flow cytometry and cell cycle phases quantified using ModFit LT software version 3.2 (Verity Software House, Topsham, Maine, USA).
Modfit lt software version 3
Manufactured by Verity Software House
Sourced in United States
ModFit LT software version 3.0 is a laboratory analysis tool designed for cell cycle analysis. The software provides capabilities for DNA content analysis and cell cycle distribution estimation.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Pi rnase staining buffer, by BD (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions)
Rnase a, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Lsrfortessa, by BD (1 mentions)
Moflo xdp flow cytometer, by Beckman Coulter (1 mentions)
Cell cycle analysis kit, by Beyotime (1 mentions)
Lsr 2, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Cellquest software version 3, by BD (1 mentions)
Facscalibur system, by BD (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Apodetect annexin 5 fitc kit, by Thermo Fisher Scientific (1 mentions)
Cyan adp flow cytometry, by Beckman Coulter (1 mentions)
Facscan, by BD (1 mentions)
Rnase a, by Beyotime (1 mentions)
Pi staining buffer, by BD (1 mentions)
19 protocols using modfit lt software version 3
1
Cell Cycle Analysis by Flow Cytometry
2
Cell cycle analysis of MDA-MB-231 cells
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MDA-MB-231 cells (with or without siRNA transfection) were plated at a density of 5 × 105 cells/dish in 60-mm dishes and incubated for 24 hours. Next, the cells were serum-deprived for 24 hours, followed by the re-addition of serum and DPPA or transfection with siRNA; the cells were then harvested 20 hours later. The cells were stained using BD PharmingenTM PI/RNase staining buffer (#550825), and the analysis was conducted using a flow cytometer (Becton Dickinson). Cell cycle modeling was performed using the Modfit LT software, version 3.2 (Verity Software House). The data were obtained from three independent experiments.
3
Cell Cycle Analysis via Flow Cytometry
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For cell cycle analysis, cells were seeded (2×105 cells/well) into 6-well culture plates and maintained overnight. Subsequently, cells were treated with Da-Ea (10, 20 or 50 µg/ml) for 24 or 48 h. Cells were washed twice with PBS. Cells were fixed in 75% ethanol at 4°C for 2 h and subsequently washed three times with PBS. Cells were stained with 500 µl PI/RNase staining buffer (BD Biosciences; Becton, Dickinson and Company) at room temperature for 15 min in dark. Following staining, cells were filtrated on a nylon membrane (pore size, 48 ìm). Cells were analysed using a FACSAriaII flow cytometer (Beckman Coulter, Inc.) and ModFit LT software, version 3.2 (Verity Software House, Inc.).
4
Cell Cycle Analysis of A549 Cells
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Briefly, A549 cells were harvested by trypsin with no EDTA (Thermo Fisher Scientific, Inc.), and washed twice with PBS. The cells were fixed with cold 70% ethanol overnight, and then stained with a solution consisting of 20 µg/ml propidium iodine (PI) (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) and 0.1% TritonX-100 (Sigma-Aldrich; Merck KGaA), 100 µg/ml RNase A (Fermentas; Thermo Fisher Scientific, Inc.) for 15 min at 37°C in the dark. Cell cycle progression was then analyzed by flow cytometry (BD LSRFortessa; BD Biosciences, Franklin Lakes, NJ, USA) and ModFit LT software (version 3.2; Verity Software House, Topsham, ME, USA).
5
Cell Cycle Analysis of EPCs
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Cell cycle analyses were performed using FCM to measure DNA content. Briefly, EPCs were harvested by centrifugation at 472 × g for 5 min at room temperature and washed with phosphate-buffered saline (PBS). Cells were fixed with 70% ethanol overnight at 4°C, washed with PBS and stained using a Cell Cycle Analysis kit (Beyotime Institute of Biotechnology, Shanghai, China) at 37°C for 30 min in the dark. Analysis was performed on ~20,000 cells/sample using a MoFlo™ XDP flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) and ModFit LT™ software version 3.2 (Verity Software House, Inc. Topsham, ME, USA).
6
Cell Cycle Analysis by DNA Content
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The analysis of the cell cycle was done by DNA content determination. Cells were plated in 6-well plates and incubated at 37° C for 24 h. GuaDex was incubated with the cells at 5 μM concentration for 24 h. The cells were harvested after incubation by a cell scraper and fixed in 4% formaldehyde in PBS at RT for at least 18 h. After fixation, cells were harvested and resuspended in 95% ethanol and kept at RT until analysis. Next, cells were washed with 2 ml dH2O, centrifuged, resuspended in 200 μL protease solution (0.1% Carlsberg’s solution Sigma protease XXIV in 0.1 M TRIS, 0.07 M NaCl, pH 7.2) and incubated at 37 °C water bath for 1 h. After incubation, 200 μL of 4′,6-diamidino-2-phenylindole (DAPI) at 8 μM solution (in 50 uM Sulphur-rhodamine 101, 1.1 M NaCl, pH 7.5) was added, and cells were analyzed on a BD LSRII (BD Biosciences, CA, USA) acquiring 10,000 events per sample. Flow cytometry data were analyzed using the ModFit LT software (version 3.2) (Verity Software House, USA).
7
Cell Cycle Analysis by Flow Cytometry
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Cells were collected for cell cycle analysis. First, cells were digested with trypsin and washed with PBS, followed by overnight fixation at 4°C with 75% ice-cold ethanol. On the second day, the fixed cells were centrifuged at 250 × g, and washed with PBS at room temperature, followed by incubation for 15 min at room temperature in the dark with 300–500 µl PI/RNase Staining Buffer (cat. no. 550825; BD Pharmingen; BD Biosciences). BD Accuri C6 flow cytometry (Becton, Dickinson and Company) was used for cell collection and the ModFit LT software version 3.2 (Verity Software House, Inc.) was used for cell cycle analysis of the samples.
8
Cell Cycle Analysis of AGS Cells with Pâté
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To analyse the AGS cell distribution in the cell cycle phases after incubation with pâté at different concentrations and times, 250,000 cells were seeded in 6 cm Petri dishes and after 48 h incubated with pâté at 50, 100 and 200 μg/mL. At the end of incubation, the cells were collected by trypsinization and stained with propidium iodide according to the method of [34 (link)]. Samples were analysed using a FACScan flow cytometer (Becton Dickinson, Milan, Italy). Quantification of cells in the different cycle phases was performed by ModFit LT software, version 3.0 (Verity Software House Inc., Topsham, ME, USA).
9
Baicalin-induced cell cycle arrest in U-2 OS cells
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U-2 OS cells (0.4×105 cells/well) were seeded on a 6-well plate, incubated for 24 h, and treated with 0, 25, 50 or 100 µM baicalin for 24 h. The cells were harvested and fixed with 70% ethanol at 4°C overnight. The fixed cells were washed twice with cold PBS and incubated for 30 min with PI/RNase at RT. The fluorescence signal was detected by the FL2 channel of a flow cytometer and the proportion of DNA at each phase was analyzed using ModFit LT software version 3.0 (Verity Software House, Inc.).
10
Cell Cycle Analysis via Flow Cytometry
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HK-2 cells were harvested, washed, and resuspended in phosphate buffered saline. Cells were then fixed with 70% ethanol and stored at 4 °C overnight. Subsequently, cells were incubated with 100 µg/mL RNase for 30 min at 37 °C and stained with 5 µg/mL propidium iodide (PI) for 10 min. Flow cytometry analyses were performed using a BD FACSCalibur (BD Biosciences). Cell cycle phase distributions were determined using Modfit LT software version 3.0 (Verity Software House, Topsham, ME, USA).
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