Anti id1 sc 488

Briefly, cells were washed with phosphate-buffered saline (PBS), harvested by trypsinization, and incubated with PE-conjugated anti-human CD133 (130-080-901, Miltenyi Biotec), APC-conjugated anti-human CD44 (559942, BD Biosciences Pharmingen, San Diego, CA, USA), and FITC-conjugated anti-human CD24 (555427, BD Biosciences Pharmingen) antibodies. For BMI-1 analysis, cells were washed with PBS, harvested by trypsinization, pre-incubated with 0.3% saponin in PBS for 10 min, and then incubated with goat anti-human BMI-1 (CAT#sc-30934, Santa Cruz Biotechnology), followed by secondary mouse anti-goat antibody (FITC conjugated). In a similar way, cells were stained with anti-Id1 (sc-488, Santa Cruz Biotechnology) and anti-BMI-1 (sc-30943, Santa Cruz Biotechnology) antibodies. Cells were resuspended in PBS and analyzed on FACSCalibur using CellQuest Pro (BD Biosciences) and FlowJo 7.6 (Tree Star, Inc., Ashland, OR, USA). Similarly, Id1⁺ and BrdU⁺ cells were analyzed with flow cytometry as above.