Multiparametric Flow Cytometry Analysis

The peripheral blood cells and splenocyte cells from various treatments were stained with anti- CD16/32, CD3, CD4, CD8, and CD19 antibodies. For thymocytic cells CD117, CD44, CD25, CD127, CD3, CD4 and CD8 were used to stain different population. Hematopoietic stem and progenitor cells were stained with lineage markers as described earlier [18 (link)]. We used following antibodies: CD117, Ly-6A/E, CD34, CD135, CD127, CD48, CD150 and CD16/32 from BD biosciences and Mac-1, Gr-1, CD4, CD8, B220, CD3, and Ter119 from Biolegend. Cells were analyzed by MACSQuant Analyzer flow cytometer (Miltenyi Biotec) and data analyzed with FlowJo software (Tree Star Inc.).

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Kapoor V., Collins A., Griffith K., Ghosh S., Wong N., Wang X., Challen G.A., Krambs J., Link D., Hallahan D.E, & Thotala D. (2020). Radiation induces iatrogenic immunosuppression by indirectly affecting hematopoiesis in bone marrow. Oncotarget, 11(19), 1681-1690.

Publication 2020

CD117 Ly-6A/E CD135 CD150 CD16/32 Ter119 MACSQuant Analyzer flow cytometer

Antibodies Cd150 Cd25 Cd44 Cells Hematopoietic Mac 1 Peripheral blood cells Progenitor cells Stem Tree

Corresponding Organization : Washington University in St. Louis

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Variable analysis

independent variables

Various treatments

dependent variables

Percentages of different cell populations (CD16/32+, CD3+, CD4+, CD8+, CD19+, CD117+, CD44+, CD25+, CD127+, CD34+, Ly-6A/E+, CD135+, CD48+, CD150+, Mac-1+, Gr-1+, B220+, Ter119+)

control variables

Peripheral blood cells
Splenocyte cells
Thymocytic cells
Hematopoietic stem and progenitor cells
Antibodies used for staining (CD16/32, CD3, CD4, CD8, CD19, CD117, CD44, CD25, CD127, CD34, Ly-6A/E, CD135, CD48, CD150, Mac-1, Gr-1, B220, CD3, Ter119)
Flow cytometer (MACSQuant Analyzer)
Data analysis software (FlowJo)

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