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. Author manuscript; available in PMC: 2020 Jul 29.
Published in final edited form as: Nature. 2020 Jan 29;578(7794):266–272. doi: 10.1038/s41586-020-1961-1

Extended Data Figure 1. Flow-sorting strategy of single basal bronchial epithelial cells.

Extended Data Figure 1

(A) Sorting of EpCam+ epithelial cells from human airway biopsies. Human hematopoietic and endothelial cells were stained with antibodies against CD45 and CD31, respectively. Within the population of cells negative for those markers, EpCam-expressing cells were gated. Single, live (DAPI-negative) cells were flow sorted from this population into individual wells of 96-well plates.

(B) qPCR analysis of clonally derived airway epithelial cell cultures. Airway basal cells express integrin alpha 6 (ITGA6), keratin 5 (KRT5), e-cadherin (CDH1) and TP63. Expression is shown in clonally derived cell cultures (n = 13 from 3 donors, coloured blue, green and orange) compared to a control bulk human bronchial epithelial cell culture expanded in the same culture conditions and a lung fibroblast cell culture that served as a negative control. Centre values and error bars indicate mean and standard error of the mean, respectively. Conditions in which no expression was detected are shown as 0.

(C) Colony-forming efficiency of CD45-/CD31-/EPCAM+ cells after single cell sorting from endobronchial biopsy samples (n = 16). For one ex-smoker, EPCAM was not used to select cells: only CD45-/CD31- cells were sorted – as expected, this is the patient with the lowest colony-forming efficiency.