Extended Data Fig. 2: FACS gating strategy. | Nature Cancer

Extended Data Fig. 2: FACS gating strategy.

From: Radiation exposure elicits a neutrophil-driven response in healthy lung tissue that enhances metastatic colonization

Extended Data Fig. 2

a-c, Example of FACS gating strategy to determine the frequency of (a) GFP+ cancer cells or (b) Ly6G+ neutrophils in the lung tissue of control/irradiated mice, or (c) Lineage–/EpCAM+ epithelial cells in the irradiated metastatic niche (Cherry-labelled, left), or unlabelled distant lung (Cherry-negative, right). All samples were gated to exclude debris and doublets, followed by live cell discrimination by DAPI staining. All gates were set based on fluorescence-minus-one (FMO) controls, containing all antibodies minus the one of interest, to determine the background signal. Importantly, lung tissue displays a high level of autofluorescence, which needs to be considered when excluding dead cells (an FMO-DAPI is critical for this).

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