Anti mouse cd29 apc

Lineage-positive cells were depleted from single-cell preparations using the EasySep Mouse Mammary Stem Cell Enrichment kit (#19757; Stemcell Technologies, Vancouver, BC, Canada). For transduced tumors, tdTomato fluorescence was used to positively select Lin− tumor cells for analysis and sorting. Single-cell preparations were resuspended (10⁷ cells/ml in HBSS+ (containing 2% fetal bovine serum with 10 mm HEPES buffer)) for antibody staining. Antibody incubations were performed on ice for 30 min. Cells were stained with anti-mouse CD24-Pacific Blue and anti-mouse CD29-APC (1:100; BioLegend, San Diego, CA, USA). Cells were either fluorescence-activated cell sorting analyzed (LSR Fortessa; BD Biosciences, Franklin Lakes, NJ, USA) or sorted (Aria II; BD Biosciences). Data analysis was performed using FlowJo, version 9.5.3 (Tree Star, Ashland, OR, USA).