Tumors were dissected from the lungs of TKO Hes1GFP/+ mice approximately 5–7 months after tumor induction and digested as previously described40 (link). The antibodies used were: CD45-PE-Cy7 (eBioscience, clone 30-F11, 1:100), CD31-PE-Cy7 (eBioscience, clone 390, 1:100), TER-119-PE-Cy7 (eBioscience, clone TER-119, 1:100), CD24-APC (eBioscience, clone M1/69, 1:200), Ncam1 (Cedarlane, clone H28-123-16, 1:100), anti-rat-IgG2a-PE (eBioscience, clone r2a-21B2, 1:200), EpCam (eBioscience, clone G8.8, 1:100), CD44-APC-Cy7 (BioLegend, clone IM7, 1:100). 1 μg/mL 7-aminoactinomycin D (Invitrogen) or DAPI was used to label dead cells.
Protocol detail
Characterization of Tumor Cell Populations
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Corresponding Organization :
Other organizations : Stanford University, OncoMed (United States), Sorbonne Université, THERANOSCAN : Biomarqueurs Théranostiques des Cancers Bronchiques Non à Petites Cellules
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Variable analysis
independent variables
- Genotype of mice (TKO Hes1GFP/+ mice)
- Characteristics of tumors dissected from the lungs
- Time after tumor induction (approximately 5–7 months)
- Digestion protocol for tumor samples
- Antibodies used for flow cytometry analysis (CD45-PE-Cy7, CD31-PE-Cy7, TER-119-PE-Cy7, CD24-APC, Ncam1, anti-rat-IgG2a-PE, EpCam, CD44-APC-Cy7)
- Dead cell staining (7-aminoactinomycin D or DAPI)
- Not explicitly mentioned
- Not explicitly mentioned
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