SVZ tissue was dissected from 300μm-thick brain sections prepared from P8 WT and CNP-hEGFR brains and processed for cell culture19. After SVZ dissection and single cell dissociation, cells were plated in 12-well cell culture dishes at a density of 50 cells/μl for 24 hours. At the time of transfection, cell cultures were approximately 60% confluent. Cell transfections were performed using the NeuroPORTER Transfection reagent (Genlantis, San Diego, CA) following manufacturer’s instructions. The cell permeable, irreversible EGFR blocker (PD168393, Calbiochem, La Jolla, CA), was used 24hrs after cell transfection. Cells were pre-incubated with PD168393 (PD) for 4hrs at 37°C in 5% CO2 and then medium was changed to fresh SCM.
