Brilliant violet 650 anti human epcam

Single-cell suspensions in FACS Buffer (HBSS [ThermoFisher 14170161, Ca/Mg-free] supplemented with 2% FCS) were pre-incubated with Fc-Block (BD 564220) before staining for surface antigens. The following antibodies were used to identify basal cells via flow cytometry: FITC anti-human THY1 (Biolegend, clone 5E10), Brilliant Violet 421 anti-human CD45 (Biolegend, clone HI30), Brilliant Violet 650 anti-human EPCAM (Biolegend, clone 9C4), APC/Cy7 anti-human ITGA6 (Biolegend, clone GoH3), PE/Cy7 anti-human NGFR (Biolegend, clone ME20.4), APC anti-human PDPN (Biolegend, clone NC-08). Cells were stained for 30 minutes on ice in FACS buffer and then washed for immediate sorting. Cells were sorted on a BD FACSAria Fusion cell sorter using BD FACSDiva software. Up to 10,000 Basal cells were sorted into 100 μL BAM banker (Wako chemicals) for Omni-ATAC-seq and cooled to −80°C using a “Mr. Frosty” freezing container (Thermo scientific). Samples were stored at −80°C until thawed for Omni-ATAC-seq. 1,000 Basal cells for bulk RNAseq were sorted directly into 5 μL TCL buffer (Qiagen). FlowJo v10 by TreeStar was used to generate plots.