Murine Lung and Blood Cell Profiling

Single-cell suspensions were prepared from perfused lungs or blood samples as described previously (Qian et al., 2011 (link)). After blocking with anti–mouse CD16/CD32 antibody (BD), the cells were stained with DAPI and fluorescent antibodies to following antigens; CD45 (30-F11), F4/80 (BM8), CD11b (M1/70), Ly6C (HK1.4), Gr1 (RB6-8C5), CD115 (AFS98), CD3 (17A2), NK1.1 (PK136; all from BioLegend); CD4 (GK1.5), B2.20 (RA3-6B2), Ly6G (1A8), CD11c (HL3; all from BD); CD8a (53–6.7), CCR5 (7A4; both from eBioscience); CCR1 (643854), CCR2 (475301; both from R&D Systems). Flow cytometry was performed using LSRII cytometer (BD) and analyzed using FlowJo software (Tree Star). In some experiments, the stained cells were sorted using FACSAria II (BD).

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Kitamura T., Qian B.Z., Soong D., Cassetta L., Noy R., Sugano G., Kato Y., Li J, & Pollard J.W. (2015). CCL2-induced chemokine cascade promotes breast cancer metastasis by enhancing retention of metastasis-associated macrophages. The Journal of Experimental Medicine, 212(7), 1043-1059.

Publication 2015

anti–mouse CD16/CD32 antibody LSRII cytometer FACSAria II

Antigens Blocking antibody Blood Ccr1 Ccr5 Cd11b Cells Dapi Flow cytometry Fluorescent antibodies Lungs Mouse Tree

Corresponding Organization :

Other organizations : University of Edinburgh, MRC Centre for Reproductive Health, Queen's Medical Centre, Albert Einstein College of Medicine

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Variable analysis

independent variables

Perfused lungs or blood samples

dependent variables

Cell populations identified by the following surface markers: CD45, F4/80, CD11b, Ly6C, Gr1, CD115, CD3, NK1.1, CD4, B2.20, Ly6G, CD11c, CD8a, CCR5, CCR1, CCR2

control variables

Procedures described in the previously published paper (Qian et al., 2011)

controls

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

Annotations

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