Sorting Cancer Cells with Flow Cytometry

For sorting of cancer cells, T8 OGO tumors were processed as previously described (Biffi et al., 2019 (link)). Cells were stained for 30 min with anti-mouse CD45-Alexa Fluor 647 (103124; BioLegend), CD326 (EpCAM)-Alexa Fluor 488 (118212; BioLegend), CD31-Alexa Fluor 647 (102416; BioLegend), and PDPN-APC/Cy7 (127418; BioLegend) and for 15 min with DAPI. DAPI/CD45/CD31/PDPN⁻ EpCAM⁺ cells were sorted on the FACSAria cell sorter (BD) and processed for PCR-based genotyping of Trp53 1loxP.

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Oni T.E., Biffi G., Baker L.A., Hao Y., Tonelli C., Somerville T.D., Deschênes A., Belleau P., Hwang C.I., Sánchez-Rivera F.J., Cox H., Brosnan E., Doshi A., Lumia R.P., Khaledi K., Park Y., Trotman L.C., Lowe S.W., Krasnitz A., Vakoc C.R, & Tuveson D.A. (2020). SOAT1 promotes mevalonate pathway dependency in pancreatic cancer. The Journal of Experimental Medicine, 217(9), e20192389.

Publication 2020

anti-mouse CD45-Alexa Fluor 647 CD326 (EpCAM)-Alexa Fluor 488 CD31-Alexa Fluor 647 PDPN-APC/Cy7 FACSAria cell sorter

Alexa fluor 488 Alexa fluor 647 Cancer Cell Dapi Epcam Mouse Trp53 Tumors

Corresponding Organization : Cold Spring Harbor Laboratory

Other organizations : Stony Brook University, University of Cambridge, Cancer Research UK, University of California, Davis, Memorial Sloan Kettering Cancer Center, Howard Hughes Medical Institute

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Variable analysis

independent variables

Sorting of cancer cells

dependent variables

DAPI/CD45/CD31/PDPN^- EpCAM^+ cells

control variables

T8 OGO tumors were processed as previously described (Biffi et al., 2019)
Cells were stained with anti-mouse CD45-Alexa Fluor 647, CD326 (EpCAM)-Alexa Fluor 488, CD31-Alexa Fluor 647, and PDPN-APC/Cy7
Cells were stained with DAPI

controls

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

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