NPCs were dissociated with accutase and then cultured for 48 hours in nonadherent plates to generate neurospheres. Neurospheres were manually picked and cultured in “Matrigel matrix (0.5 mg Matrigel was plated in cold NPC media on a 96-well plate 1 hour prior to neurosphere plating; following neurosphere picking, an additional 0.5 mg Matrigel was added in cold NPC media per 96-well plate). DAPI-stained neurospheres were imaged at 48 hours. Average radial migration from each neurosphere was measured using NIH ImageJ40,74.
