Diva v8.0.1

Manufactured by BD

The DIVA (v8.0.1) is a flow cytometry software developed by BD. It is designed to analyze and process data from flow cytometry experiments. The software provides tools for data acquisition, visualization, and analysis.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (1 mentions) Fix and perm cell fixation and cell permeabilization kit, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 conjugate, by Cell Signaling Technology (1 mentions) Anti human cd90 pe, by BioLegend (1 mentions) Kdm4b, by Abcam (1 mentions) Pe anti human cd34 antibody, by BioLegend (1 mentions) Macsquant analyzer vyb, by Miltenyi Biotec (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

2 protocols using diva v8.0.1

Quantifying CM Proliferation via EdU Incorporation

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FACS quantification of rates of EdU incorporation was carried out using littermate Dot1L cKOs and Ctrls. EdU detection was done in cell suspensions using the Click-iT® EdU Alexa 647 kit (Thermo Fisher Scientific; C10340), according to the manufacturer’s instructions. TdTomato signal was used to discriminate CMs from other lineages of cardiac cells. Stained cells were analyzed using a FACS Canto II flow cytometer (BD Bioscience). DIVA (v8.0.1) and FlowJo (v10.8.1) software (BD Pharmingen) were used for data acquisition and analysis.

Cattaneo P., Hayes M.G., Baumgarten N., Hecker D., Peruzzo S., Aslan G.S., Kunderfranco P., Larcher V., Zhang L., Contu R., Fonseca G., Spinozzi S., Chen J., Condorelli G., Dimmeler S., Schulz M.H., Heinz S., Guimarães-Camboa N, & Evans S.M. (2022). DOT1L regulates chamber-specific transcriptional networks during cardiogenesis and mediates postnatal cell cycle withdrawal. Nature Communications, 13, 7444.

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Multicolor Flow Cytometry Characterization

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Cells were analyzed with an LSR II flow cytometer (BD Biosciences) using Diva v8.0.1 (BD Biosciences) or a Miltenyi MACSQuant VYB analyzer. Cell permeabilization was performed using the Fix and Perm Cell Fixation and Cell Permeabilization Kit (ThermoFisher Scientific, GAS003) following the manufacturer’s instructions. Primary antibodies used were NANOG (D73G4) XP® Rabbit mAb (1:100) (Alexa Fluor® 647 Conjugate, Cell signaling 5448), THY-1 (1:100) (PE anti-human CD90 (Biolegend 328110)), KDM4B (1:100) (Abcam ab191434), CD73 (1:100) (APC/Cy7 anti-human CD73 Antibody (Biolegend 344021)), CD105 (1:100) (APC anti-human CD105 Antibody (Biolegend 323207), (1:100) CD146 (APC anti-human CD146 (1:100) (Biolegend 361015)), CD144 (1:100) (PE anti-human CD144 Antibody (Biolegend 348505), CD34 (1:100) (PE anti-human CD34 Antibody (Biolegend 343605).

Stefano B.D., Luo E.C., Haggerty C., Aigner S., Charlton J., Brumbaugh J., Ji F., Jiménez I.R., Clowers K.J., Huebner A.J., Clement K., Lipchina I., de Kort M.A., Anselmo A., Pulice J., Gerli M.F., Gu H., Gygi S.P., Sadreyev R.I., Meissner A., Yeo G.W, & Hochedlinger K. (2019). The RNA helicase DDX6 controls cellular plasticity by modulating P-body homeostasis. Cell stem cell, 25(5), 622-638.e13.

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