Characterizing Regulatory T Cells by FACS

MLN single cell suspensions were stained and analyzed according to (42 (link)). Cells were stained for: viability (BV510), CD4 (BUV395), and Foxp3 (BV421), all purchased from BD Biosciences, Canada. Cells were blocked with rat α-mouse CD16/32 from Biolegend, USA, surface stained for CD4 and stained intracellularly for Foxp3. Cells were run on an LSRFortessa X-20 flow cytometer and data were analyzed using FlowJo software (v10 & 7.6.5, FlowJo LLC, USA). For analysis, doublets and dead cells were removed from the analysis.

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Ariyaratne A., Kim S.Y., Pollo S.M., Perera S., Liu H., Nguyen W.N., Leon Coria A., de Cassia Luzzi M., Bowron J., Szabo E.K., Patel K.D., Wasmuth J.D., Nair M.G, & Finney C.A. (2022). Trickle infection with Heligmosomoides polygyrus results in decreased worm burdens but increased intestinal inflammation and scarring. Frontiers in Immunology, 13, 1020056.

Publication 2022

LSRFortessa X-20 flow cytometer

Cells Mouse X flow

Corresponding Organization : University of Calgary

Other organizations : University of California, Riverside

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Variable analysis

independent variables

Staining for viability (BV510), CD4 (BUV395), and Foxp3 (BV421)

dependent variables

Cell population analysis

control variables

Blocking with rat α-mouse CD16/32
Gating of doublets and dead cells
Use of an LSRFortessa X-20 flow cytometer
Data analysis using FlowJo software (v10 & 7.6.5)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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