Characterizing Mouse Blood and Bone Marrow

For mouse phenotyping peripheral blood and bone marrow from femurs were collected from animals of different genotypes and processed as previously described.^{16 (link)} For flow analysis, samples were stained with Gr1-PE (Miltenyi Biotec, Woking, UK), CD11b (Mac1)-FITC (Becton Dickinson), CD71-PE (Becton Dickinson), Ter119-Fitc (Becton Dickinson). For analysis of progenitor populations BM cells were lysed with Red Blood Cell buffer (NH4, Cl) and then enriched with the EasySep Hematopoietic Progenitor Cell Isolation Kit (StemCell Technologies, Cambridge, UK) followed by staining with following antibodies Sca1-PB, c-KIT-Acy7, CD34-FITC, Flk2-PeCy5, Cd16/32-PE, CD150-PeCy7, CD48-APC.

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Mupo A., Seiler M., Sathiaseelan V., Pance A., Yang Y., Agrawal A.A., Iorio F., Bautista R., Pacharne S., Tzelepis K., Manes N., Wright P., Papaemmanuil E., Kent D.G., Campbell P.C., Buonamici S., Bolli N, & Vassiliou G.S. (2016). Hemopoietic-specific Sf3b1-K700E knock-in mice display the splicing defect seen in human MDS but develop anemia without ring sideroblasts. Leukemia, 31(3), 720-727.

Publication 2016

Gr1-PE CD11b (Mac1)-FITC CD71-PE Ter119-Fitc EasySep Hematopoietic Progenitor Cell Isolation Kit CD34-FITC Cd16/32-PE

Animals Antibodies Blood Bone marrow Buffer C kit Cd11b Cd150 Cd71 Femurs Fitc Genotypes Hematopoietic progenitor cell Isolation Mouse Populations Red blood cell Sca1 Stemcell

Corresponding Organization : University of Cambridge

Other organizations : Wellcome/MRC Cambridge Stem Cell Institute, European Bioinformatics Institute, Memorial Sloan Kettering Cancer Center

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Variable analysis

independent variables

Genotypes of animals

dependent variables

Peripheral blood
Bone marrow from femurs
Populations of hematopoietic progenitor cells (Sca1, c-KIT, CD34, Flk2, Cd16/32, CD150, CD48)

control variables

Sample processing method as previously described
Antibodies used for flow cytometry analysis (Gr1-PE, CD11b (Mac1)-FITC, CD71-PE, Ter119-Fitc, Sca1-PB, c-KIT-Acy7, CD34-FITC, Flk2-PeCy5, Cd16/32-PE, CD150-PeCy7, CD48-APC)

controls

No positive or negative controls were explicitly mentioned.

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