Transient transfections and Luciferase assays were performed in 60% confluent SVZ cell cultures (CNP-hEGFR and WT) using NeuroPORTER as described above, and 1.5μg of PGL3 basic, wt phEGFR (Millipore), pCBF1-Luciferase reporter plasmid (gift of Dr. Gabriel Corfas, Harvard Medical School) or Hes1 Luciferase reporter plasmid, and Dll expression plasmid (gift of Dr. Alanis Israel, Unite de Biologie Moleculaire de l’Expression Genique, Centre National de la Recherche Scientifique) reporter plasmids and 0.3μg of expression vectors. Notch expression vectors used were a kind gift of Dr. Raphael Kopan (Department of Molecular Biology and Pharmacology, Washington University). Luciferase assays were performed 48hr after transfection using the Dual Assay Luciferase kit (Promega). Cotransfected TK–renilla Luciferase was used to normalize samples for transfection efficiency and for sample handling. Cells were lysed, and luciferase activity was measured following the protocol recommended by the manufacturer.
