Mouse lung single-cell flow cytometry

Mouse lung single-cell suspensions and the procedures of staining the cells for flow cytometry and data analysis were described previously (Barkauskas et al., 2013 (link); Chen et al., 2012 (link); Liang et al., 2016 (link)). Flow cytometry was performed using a LSRFortessa flow cytometer and FACSAria III sorter (BD, San Jose, CA) and the data were analyzed using Flow Jo 9.9.6 software (Tree Star, Ashland, OR). Primary antibodies EpCAM-PE-Cy7 (clone G8.8, Catalog # 118216, RRID AB_1236471) were from BioLegend. CD24-PE (clone M1/69, Catalog # 12-0242-82, RRID AB_467169), Sca-1 (Ly-6A/E)-APC (clone D7, Catalog # 17-5981-82, RRID AB_469487), CD31 (PECAM-1) (clone 390, Catalog # 13-0311-85, RRID AB_466421), CD34 (clone RAM34, Catalog # 13-0341-85, RRID AB_466425), and CD45 (clone 30-F11, Catalog # 13-0451-85, RRID AB_466447) were all from eBioscience (San Diego, CA). Streptavidin-APC-Cy7 (catalog # 405208) was from BioLegend (San Diego, CA).

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Liang J., Huang G., Liu X., Liu N., Taghavifar F., Dai K., Yao C., Deng N., Wang Y., Chen P., Hogaboam C., Stripp B.R., Parks W.C., Noble P.W, & Jiang D. (2023). Reciprocal interactions between alveolar progenitor dysfunction and aging promote lung fibrosis. eLife, 12, e85415.

Publication 2023

LSRFortessa flow cytometer FACSAria III sorter EpCAM-PE-Cy7 clone G8.8, CD24-PE (clone M1/69, CD34 (clone RAM34, CD45 (clone 30-F11, Streptavidin-APC-Cy7

Antibodies Clone Epcam Flow cytometry Lung Mouse Pecam 1 Sca 1 Streptavidin Tree

Corresponding Organization :

Other organizations : Cedars-Sinai Medical Center, Lung Institute

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Variable analysis

independent variables

Mouse lung single-cell suspensions

dependent variables

Flow cytometry data

control variables

Staining procedures for flow cytometry
Flow cytometry performed using LSRFortessa flow cytometer and FACSAria III sorter
Data analyzed using Flow Jo 9.9.6 software

controls

Positive control: Primary antibodies EpCAM-PE-Cy7, CD24-PE, Sca-1 (Ly-6A/E)-APC, CD31 (PECAM-1), CD34, and CD45
Negative control: Streptavidin-APC-Cy7

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