Pe cy7 strep

Before flow cytometry analysis or sorting, cells were dissociated as described above or washed and dissociated using TrypLE (Gibco, Carlsbad, CA, USA). Cells were resuspended in a 100μL staining volume of fluorescence activated cell sorting (FACS) buffer (Hanks' balanced salt solution + 0.1% bovine serum albumin) and kept on ice. The following antibodies were used: CD133/1, and CD133/2 biotin (Miltenyi Biotech, San Diego, CA, USA), CD90 (Biolegend, San Diego, CA, USA), CD15 (BD Bioscience, San Jose, CA, USA), CD49f (BD Bioscience, San Jose, CA, USA), AB25 (Miltenyi Biotech, San Diego, CA, USA) and PE-CY7 strep (BD Biosciences, San Jose, CA, USA). All analyses were conducted using a LSR II FACS machine (BD Biosciences, San Jose, CA, USA) or sorted on an ARIA-II (BD Biosciences, San Jose, CA, USA) at the Stanford University FACS facility. Appropriate isotype controls were used to control for nonspecific isotype background.