Figure 2. Gating strategy to purify basal, luminal and immune cell subsets.

FACS plots demonstrate the gating strategy used to isolate total cells based on forward scatter (FSC) and side scatter (SSC), and to exclude doublets using SSC width vs. area. Total cells can be analyzed by expression of CD49f and Trop2, as has been previously reported. A more complex sorting strategy includes separation of immune cells (CD45⁺) and epithelial cells (Trop2⁺). The CD45+ fraction is comprised of both CD8+ lymphocytes and CD8- immune cells, while the Trop2+ epithelium can be subdivided into basal (Trop2⁺CD49f^hiCD24^lo/+) and luminal (Trop2⁺CD49f^loCD24⁺) fractions.