Annexin V and Propidium Iodide Staining

Thymocytes were isolated following the procedure described previously^{3 (link)}. Irradiation was conducted about 1 h after cell culture in vitro. At 24 h after irradiation, thymocytes were collected, washed using PBS supplemented with Ca²⁺ and Mg²⁺, and then stained with a FITC-conjugated annexin V (BD Pharmigen) and propidium iodide (5 μg/ml, Sigma-Aldrich). Data were collected from at least 20,000 cells by FACSCanto (BD Pharmingen) and analyzed by Flowjo (Tree Star, Inc) without knowledge of the genotype or treatment by a single observer (C-LL).

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Hasapis S., Caraballo I., Sears T.J., Brock K.D., Cart J.B., Moding E.J, & Lee C.L. (2023). Characterizing the role of Phlda3 in the development of acute toxicity and malignant transformation of hematopoietic cells induced by total-body irradiation in mice. Scientific Reports, 13, 12916.

Publication 2023

FITC-conjugated annexin V propidium iodide FACSCanto

Annexin v fitc Cell culture Cells Genotype Irradiation Propidium iodide Thymocytes Tree

Corresponding Organization :

Other organizations : Duke University Hospital, Duke Medical Center, Stanford University, Duke University

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Variable analysis

independent variables

Irradiation

dependent variables

Annexin V staining
Propidium iodide staining

control variables

Thymocytes isolated following the procedure described previously
Cells washed using PBS supplemented with Ca2+ and Mg2+
Data collected from at least 20,000 cells by FACSCanto and analyzed by Flowjo without knowledge of the genotype or treatment by a single observer (C-LL)

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