Journal: Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.]

Article Title: Assaying cell cycle status using flow cytometry

doi: 10.1002/0471142727.mb2806s111

Figure Lengend Snippet: Lists of fluorophores for dye dilution proliferation assay

Article Snippet: Laser Manufacturer CFSE 495 519 488 nm Various CellTrace TM Violet 405 450 405 nm Life Technologies BD HorizonTM Violet Cell Proliferation Dye (VPD450) 404 448 405 nm BD Biosciences Cell Proliferation Dye eFluor®450 405 450 405 nm eBioscience CytoTrack TM Blue 403 454 405 nm BIO-RAD Oregon Green 488 carboxylic acid diacetate, SE (carboxy-DFFDA SE) 496 524 488 nm Life Technologies SNARF-1 carboxylic acid, acetate, SE 514 586 488 nm Life Technologies CytoTrack TM Green 511 525 488 nm BIO-RAD CytoTrack TM Yellow 542 556 532 nm BIO-RAD CellTrace TM Far Red DDAO-SE 647 657 633 nm Life Technologies Cell Proliferation Dye eFluor®670 647 670 633 nm eBioscience CytoTrack TM Red 628 643 633 nm or 640 nm BIO-RAD Open in a separate window Lists of fluorophores for dye dilution proliferation assay Critical Parameters Proper cell density Cell density should be optimized because confluent cultures may cause growth arrest by contact inhibition, which leads to G0/G1 arrest of the cell cycle.

Techniques:

Journal: Scientific Reports

Article Title: Hypoxic Stress Decreases c-Myc Protein Stability in Cardiac Progenitor Cells Inducing Quiescence and Compromising Their Proliferative and Vasculogenic Potential

doi: 10.1038/s41598-017-09813-x

Figure Lengend Snippet: Inhibition of c-Myc decreases cardiac progenitor cell proliferation and induces quiescence. ( A ) Representative histogram showing CPC proliferation analysis by CellTrace dye dilution 72 hours after treatment with the c-Myc inhibitor 10058-F4 (dotted line) and DMSO control (solid line, light gray) compared to baseline (solid line, dark gray). Graph indicates the CellTrace mean fluorescence intensity showing less dye dilution in samples treated with the c-Myc inhibitor relative to DMSO control, an indication of delayed growth (n = 4, *p < 0.03). ( B ) Quantification of Ki67 expression 72 hours after treatment of CPCs with c-Myc inhibitor and DMSO control by fluorescence activated cell sorting. Graph indicates the average of Ki67 mean fluorescence intensity (MFI) in control (black bars) and c-Myc inhibitor-treated CPCs (gray bars) (n = 4, *p < 0.03). ( C ) Cell cycle analysis of CPCs after c-Myc inhibition. Dot Plots show representative images of cells stained with 7-AAD and Ki67 and analyzed 72 hours later. Graph indicates the average percentage of cells in each phase of the cell cycle in DMSO control (black bars) and c-Myc inhibitor treated CPCs (gray bars) (n = 4, *p < 0.05).

Article Snippet: The effect of c-Myc inhibition on CPC growth was determined by staining cells with 1 μM of CellTrace Far Red Cell Proliferation Dye (Thermo Fisher Scientific) followed by treatment with 40 μM the c-Myc inhibitor 10058-F4 (Sigma-Aldrich, #F3680) or vehicle control (DMSO).

Techniques: Inhibition, Control, Fluorescence, Expressing, FACS, Cell Cycle Assay, Staining

Journal:

Article Title: Nap1l2 Promotes Histone Acetylation Activity during Neuronal Differentiation ▿

doi: 10.1128/MCB.00789-07

Figure Lengend Snippet: Maintenance, proliferation, and apoptosis of neural precursors from which Nap1l2 was deleted. (A) The percentages of Sox1-GFP-positive cells in suspension culture were analyzed by FACS. The data summarize three independent experiments using three different 46CΔNap1l2 ES cell clones. Error bars represent standard errors of the means. (B) GFP expression in embryoid bodies of 46CloxPNap1l2loxP (WT) and 46CΔNap1l2 cells (D) at day 21 is shown. (C) Sox1-GFP-positive cells were labeled using CellTrace Far Red DDAO-SE at day 10 of suspension culture, and the 24-h Far Red (FR) kinetics was analyzed by FACS. Results for wild-type cells are represented by dark lines, and results for mutant cells are represented by light-gray lines. Data are representative of two experiments. (D) Relative quantification in arbitrary units (AU) and standard deviations of Par-4 expression by real-time PCR (bars) and determination of percentages of annexin V-positive cells by FACS (lines), given as the means ± standard errors of the means of two independent experiments, are shown for 46CloxPNap1l2loxP (dark bars and solid line) and 46CΔNap1l2 cells (light bars and dashed line) at different stages of neuronal differentiation. NS, neural stem cell; N, neuron.

Article Snippet: Cell tracer labeling was done using CellTrace Far Red DDAO-SE (Molecular Probes) at 25 μM in PBS-1% bovine serum albumin (BSA) for 10 min at 37°C.

Techniques: Suspension, Clone Assay, Expressing, Labeling, Mutagenesis, Quantitative Proteomics, Real-time Polymerase Chain Reaction

Journal: The FASEB Journal

Article Title: A human co‐culture cell model incorporating microglia supports glioblastoma growth and migration, and confers resistance to cytotoxics

doi: 10.1096/fj.201901858rr

Figure Lengend Snippet: FIGURE 1 Labeling and viability of GBM and MG cells marked with CellTrace. A, Percentage of CFSE+ U-87 MG, SNB-19, and UP-007 cells along 9 days of culture. Percentage of CFSE-labeled cells is given compared to an unstained control. B, Viability of U-87 MG, SNB-19, and UP-007 cells stained with CellTrace CFSE (5 μM) compared to a control of unstained cells. C, Percentage of Far Red+ CHME3 cells along 9 days of culture. D, Viability of CHME3 stained with CellTrace (1 μM) when compared to a control of unstained cells. E, Histogram of the frequency of distributions of the labeling of CFSE+ GBM cells showing a decrease of fluorescence intensity along the days of culture even though the whole cell population remains positive for CFSE. Mean ± SEM (N = 3, mean of three independent experiments). P > .05 when comparing cell viability of GBM or CHME-3 labeled cells (CFSE and Far Red, respectively) to unlabeled cells

Article Snippet: Cell labeling optimization screening was performed with all GBM cells and MG using the CellTrace carboxyfluorescein succinimidyl ester (CFSE) and a CellTrace Far Red (Invitrogen, Fisher Scientific, Loughborough, UK), respectively.

Techniques: Labeling, Control, Staining, Fluorescence