Analyzing Mouse Hematopoietic Cell Subsets

Bone marrow (BM) cells isolated from the femur and tibia were prepared as described previously (Lei et al., 2018 (link)). All collected cells were treated with ACK (red blood cell lysis) buffer before analysis. To analyze hematopoietic cells, monoclonal antibodies from BD Biosciences (San Jose, CA, United States), eBioscience (San Diego, CA, United States), and BioLegend (San Diego, CA, United States) recognizing the following surface markers were used: Sca-1 (D7), c-Kit (2B8), CD150 (mShad150), CD48 (HM48-1), CD127 (A7R34), CD16/32 (93), Gr-1 (RB6-8C5), Mac-1 (M1/70), B220 (RA3-6B2), CD3e (145-2C11), CD45.1 (A20), CD45.2 (104), Ki67 (7B11), and streptavidin. The mouse lineage cocktail contained anti-CD3, Mac-1, Gr-1, B220, Ter-119 (TER-119), and NK1.1 (PK136) antibodies (eBioscience). Cell analysis was performed using a FACS LSR Fortessa (BD Biosciences) and cell sorting was performed using a FACS AriaIII (BD Biosciences). Data were analyzed using FlowJo10.0 software (TreeStar, Ashland, OR, United States).

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Liu Y., Chen Y., Deng X, & Zhou J. (2020). ATF3 Prevents Stress-Induced Hematopoietic Stem Cell Exhaustion. Frontiers in Cell and Developmental Biology, 8, 585771.

Publication 2020

NK1.1 (PK136) FACS LSR Fortessa FACS AriaIII

Anti cd3 Antibodies Bone marrow cells Buffer C kit Cd150 Cell Femur Hematopoietic Mac 1 Monoclonal antibodies Mouse Red blood cell Sca 1 Streptavidin Tibia

Corresponding Organization : Tianjin Medical University

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Variable analysis

independent variables

Preparation of bone marrow (BM) cells from the femur and tibia as described previously (Lei et al., 2018)

dependent variables

Analysis of hematopoietic cells using monoclonal antibodies recognizing surface markers: Sca-1, c-Kit, CD150, CD48, CD127, CD16/32, Gr-1, Mac-1, B220, CD3e, CD45.1, CD45.2, Ki67, and streptavidin
Cell analysis performed using a FACS LSR Fortessa
Cell sorting performed using a FACS AriaIII

control variables

Treatment of all collected cells with ACK (red blood cell lysis) buffer before analysis
Mouse lineage cocktail containing anti-CD3, Mac-1, Gr-1, B220, Ter-119, and NK1.1 antibodies (eBioscience)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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