“…For in vitro experiments, on serial time points post-TMZ or DMSO addition (days 2, 4, 6, and 8) cells were collected, labeled, and analyzed for both surface and intracellular markers as previously described. 43 The following human-specific antibodies were used: anti-CD133-PE (2 : 100; Miltenyi Biotec, Auburn, CA, USA), anti-CD15-APC (5 : 100; BD Pharminogen, San Diego, CA, USA), anti-Oct4-FITC (5 : 100; Millipore, Billerica, MA, USA), antiKi67-FITC (4 : 100; Abcam, Cambridge, MA, USA), biotinylated anti-SOX2 (4 : 100; R&D Systems), purified rabbit anti-Nestin (4 : 100; Covance, Berkeley, CA, USA), purified monoclonal anti-HIF1A (hypoxia-inducible factor-1A; 4 : 100; Abcam), purified monoclonal anti-HIF2A (4 : 100; Millipore), purified polyclonal rabbit anti-MGMT (4 : 100; Cell Signaling, Beverly, MA, USA); and anti-mouse CD45 eFluor450 (eBioscience, San Diego, CA, USA). In addition, we used as secondary antibodies anti-rabbit IgGk1 FITC (1 : 500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-mouse IgGk1 PB (1 : 500; Abcam), and Streptavidin PB (1 : 500; Molecular Probes, Eugene, OR, USA).…”