NPCs were dissociated with Accutase and then cultured for 72 h in non-adherent plates to generate neurospheres. Neurospheres were manually picked and cultured in Matrigel matrix (0.5 mg Matrigel was plated in cold NPC media to a 96-well plate 1 h before neurosphere plating; following neurosphere picking, additional Matrigel was added, again as 0.5 mg in cold NPC media per 96-well plate). Average migration from each neurosphere was measured using NIH Image J.
