Chunk #4 — METHODS — Immunohistochemistry and antibodies

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Notch and EGFR pathway interaction regulates neural stem cell number and self-renewal.

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Freshly cut, floating tissue sections (20–40μm) from P8 and P90 mice were prepared as previously described14,15. Primary antibody dilutions were: 0.5μg/ml for the specific anti-hEGFR (Biofluids, Caramillo, CA); 1:500 for anti-BrdU (Accurate, Westbury, NY), anti-NG2 antibody (Chemicon, Temecula, CA), monoclonal anti-GFAP (mouse monoclonal, Sigma Aldrich, St. Louis, MO), polyclonal anti-GFAP (rabbit polyclonal, Covance, Berkeley, CA), anti-S100β (DAKO, Denmark; rabbit anti-human clone A5110), and anti-Nestin (Chemicon); 1:50 for anti-LeX (MMA clone, BD Biosciences, San Jose, CA), anti-full length Notch-1, Jagged1, Delta like-1 (Dll) and anti-NICD (from Developmental Studies Hybridoma Bank, Iowa, IA).