Proliferative NSPCs were treated with either 1 or 10 μM UK5099 (no. PZ0160, Sigma-Aldrich) for 72 hours. For quiescence experiments, NSPCs were treated with 1 or 10 μM UK5099 (no. PZ0160, Sigma-Aldrich), 1 μM lactate dehydrogenase A inhibitor (GSK 2837808A, Tocris, 5189/10), and 1, 2.5, or 5 mM aspartate (no. 1690.2, Roth) simultaneously with quiescence induction. For exit experiments, quiescent NSPCs were treated with UK5099 after establishment of quiescence.
For differentiation experiments, proliferative NSPCs were treated with 1 or 10 μM UK5099 for 48 hours. Then, cells were washed off and plated (65,000 cells/cm2) on coated coverslips in media containing one-fifth of the growth factors and indicated UK5099 concentration. Similar to UK5099 treatment, 1, 2.5, or 5 mM aspartate (no. 1690.2, Roth) was used during the induction of quiescence.
For differentiation experiments, proliferative NSPCs were treated with 1 or 10 μM UK5099 for 48 hours. Then, cells were washed off and plated (65,000 cells/cm2) on coated coverslips in media containing one-fifth of the growth factors and indicated UK5099 concentration. Similar to UK5099 treatment, 1, 2.5, or 5 mM aspartate (no. 1690.2, Roth) was used during the induction of quiescence.