Subconfluent monolayers were harvested by trypsinization, washed, and fixed in ice-cold 70% ethanol, added dropwise while vortexing. The samples were stored at 4°C until use. Prior to analysis, the samples were washed in phosphate buffered saline (PBS; Sigma-Aldrich), resuspended in 1 mg/mL RNase (Sigma-Aldrich) and 50 μg/μL propidium iodide (PI; Sigma-Aldrich), and incubated for 30 min at room temperature prior to analysis on a FACS Accuri C6 (Accuri Cytometers Inc., Ann Arbor, MI). Lymphocytes were used as an internal diploid control and prepared for DNA analysis as described above. Cell cycle data were analyzed in ModFit LT 3.3 (Verity Software House, Topsham, ME). DNA Index (DI), which measures variations in ploidy, was calculated by the following formula: Mean G1 DNA fluorescence of cell sample/mean G1 DNA fluorescence of lymphocytes. Aneuploid cells are defined as cells which differ with more than 10% from a diploid DI = 1.0.
Modfit lt 3
Manufactured by Verity Software House
Sourced in United States, China, Montenegro
ModFit LT 3.2 is a software application developed by Verity Software House for the analysis of flow cytometry data. It provides tools for cell cycle analysis, DNA and protein quantification, and multiparameter data visualization.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (20 mentions)
Facscalibur flow cytometer, by BD (18 mentions)
Propidium iodide, by Merck Group (15 mentions)
Rnase a, by Merck Group (14 mentions)
Facsdiva software, by BD (11 mentions)
Flow cytometry, by BD (9 mentions)
Facscan, by BD (9 mentions)
Lsrii flow cytometer system, by BD (9 mentions)
Pi rnase staining buffer, by BD (8 mentions)
Cell cycle and apoptosis analysis kit, by Beyotime (7 mentions)
Facscanto 2, by BD (7 mentions)
Facscan flow cytometer, by BD (6 mentions)
Facscanto 2 flow cytometer, by BD (5 mentions)
Cell cycle staining kit, by MultiSciences Biotech (4 mentions)
Cellquest software, by BD (4 mentions)
Cytomics fc 500 mpl, by Beckman Coulter (4 mentions)
Facsaria, by BD (3 mentions)
Propidium iodide, by BD (3 mentions)
Cell cycle detection kit, by Keygen Biotech (3 mentions)
Cycletest plus dna reagent kit, by BD (3 mentions)
207 protocols using modfit lt 3
1
Cell Cycle Analysis by Flow Cytometry
2
Cell Cycle Analysis of M14 Cells Treated with UDCA
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A total of 3×105 M14 cells/well were seeded onto 6-well plates and allowed to adhere at 37°C and 5% CO2 for 12 h and following treatment with different concentrations of UDCA (0, 100, 200, and 300 µg/ml) for 48 h, cells were collected by centrifuging at 1,500 × g at 4°C for 10 min and the precipitations were washed once with PBS. The cells were fixed with a pre-cooled ethanol-PBS mixture (7:3, v/v) at 4°C for 60 min and washed twice with PBS. Finally, 10 mg/ml RNase and 1 mg/ml PI was added followed by incubating at 37°C for 30 min in the dark. Following filtration of the cells with a 300-mesh sieve, the cells were detected by flow cytometry and processed using the ModFit LT 3.3 (Verity Software House, Inc., Topsham, ME, USA) analysis software.
3
Cell Cycle Analysis of Myc Inhibitors
4
Cell Cycle Analysis of Myc Inhibitors
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Sub-confluent cultures of cells at >90% viability were plated into fresh medium at densities that were typically about 10% of those attained at the point of density arrest. The next day, Myc inhibitors at the stated concentrations were added for 24-48 hr. The cells were then harvested, washed twice in PBS and stained with propidium iodide as previously described [29 (link), 83 (link)]. Cell cycle analyses were performed on a BD FACSCalibur™ flow cytometer (Becton Dickinson, Inc. Franklin Lakes, NJ) and analyzed using ModFit LT 3.3 (Verity Software House, Topsham, ME).
5
Flow Cytometry Analysis of Brain MNCs
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Brain MNCs were cultured as described (32 (link)). Cells were collected and stained with the PI/RNAs staining following the manufacturer's instructions (Cell Signaling Technology, Danvers, MA, USA). The data were acquired by flow cytometry and analyzed with ModFit LT 3.3 (Verity Software House, Topsham, ME) after debris and doublets were gated out.
6
Apoptosis and Cell Cycle Analysis
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Cell apoptosis was analyzed using flow cytometry. Briefly, the cell suspensions were washed with PBS and cell density was adjusted so that each well contained 5 × 106 cells, and the cells were digested with trypsin and centrifuged at 1,000 rpm at room temperature for 5 min. T24 cells were stained with 100 µl of propidium iodide (PI) (P4170, Merck, Germany) and annexin V FITC (556,419, BD, USA) binding buffer and incubated for 15 min at normal temperature in the absence of light. Apoptosis was measured by a BD FACSAria III system (Beijing Jiamay Biotech) and analyzed with Modfit LT3.3 (Verity Software House, Topsham, ME, USA). For cell cycle analysis, cells were fixed and stained for 30 min with PI staining buffer containing 50 µg/mL PI and 100 µg/mL RNase A and analyzed by BD FACSAria III flow cytometer.
7
Cell Cycle Analysis of Anticancer Compounds
8
Cell Cycle Analysis by Flow Cytometry
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Cells were synchronised with 2 mM Thymidine (Sigma-Aldrich, USA), and released into fresh media. Cells (and floaters) were harvested and fixed in 100% ethanol overnight. Fixed cells were treated with 50 μg/ml RNase and stained with propidium iodide. Cell cycle profiles were analysed using a BD Accuri Flow Cytometer (Beckman Coulter, Fullerton, CA, USA). Quantification of the percentage of cells at different cell cycle stages was performed using the ModFit LT 3.3 software (Verity Software House, USA).
9
Cell Cycle Analysis of siPCDH18 Transfected Cells
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Following 48 h of incubation, NCM460 cells (1 × 106) transfected with siPCDH18 and siNC were harvested and wash twice wish PBS, fixed with ice-cold 70% ethanol overnight at 4 °C. Then, the cells were treated with 20 mg/l propidium iodide (PI) (Thermo Scientific Inc. cat#P1304MP) at 4 °C for 30 min in the dark and sorted by Beckman coulter cell Lab Quanta SC. The cell phase distribution was analysed by ModFit LT 3.1 software (Verity Software House, USA). Each sample was tested in duplicate.
10
Cell Viability and Cell Cycle Analysis of siIGF2BP3 Knockdown
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The cell viability assay was performed 24 h after transfection of siNC and siIGF2BP3 with a CCK-8 kit from Beyotime (Beijing, China). After transfection, cells were plated in 96-well dishes at a concentration of 5 × 103 cells/well and cultured in DMEM containing 15% FBS for cell attachment. Cell viability was measured with CCK-8 reagent following the manufacturer’s protocol at the indicated time points (24, 48 and 72 h).
Cell cycle analysis was performed 48 h after transfection of siNC and siIGF2BP3. Cells were washed twice with ice-cold PBS, harvested, and fixed with 70% ethanol at 4°C overnight. Then, the cells were stained with a Cell Cycle and Apoptosis Analysis Kit (Beyotime, Beijing, China) at 37°C for 30 minutes and detected by flow cytometry (Becton-Dickinson, San Jose, CA, USA). Cell cycle distributions were analysed with ModFit LT 3.1 software (verity Software House, Inc., Topsham, ME, USA).
Cell cycle analysis was performed 48 h after transfection of siNC and siIGF2BP3. Cells were washed twice with ice-cold PBS, harvested, and fixed with 70% ethanol at 4°C overnight. Then, the cells were stained with a Cell Cycle and Apoptosis Analysis Kit (Beyotime, Beijing, China) at 37°C for 30 minutes and detected by flow cytometry (Becton-Dickinson, San Jose, CA, USA). Cell cycle distributions were analysed with ModFit LT 3.1 software (verity Software House, Inc., Topsham, ME, USA).
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