Effects of Chase ABC on the differentiation of NPCs in vitro. (A-D) Treatment of Chase ABC on NPCs for 5 days resulted in a substantial increase in the number of β-III tubulin+ neurons (green, B) and decrease in the number of GFAP+ astrocytes (green, D) as compared to the control (A and C, respectively). (E, F) No significant difference was found in the number of O4+ (red) between the two groups. However, in the Chase ABC treated group, the O4+ oligodendrocytes showed more mature morphology (F). (G, H) Chase ABC treatment brought about more mature MBP+ oligodendrocytes (green, H) as compared to the control in differentiated NPCs. Hoechst 33342 was used as a nuclear marker for the estimation of cell numbers (A-H). Scale bar: 50 μm. (I) Quantitative data demonstrate that treatment of NPCs with Chase ABC resulted in a significant increase in the number of β-III tubulin+ neurons (P < 0.05), decrease of GFAP+ astrocytes (P < 0.01), no change of O4+ oligodendrocytes (P > 0.05) and increase of MBP+ oligodendrocytes (P < 0.01). Data were shown as mean ± SEM of three independent experiments with a total of more than 1000 cells examined in each group. * P < 0.05, ** P < 0.01. (J) Western blot analysis shows that Chase ABC treatment resulted in a marked increase in the expression of β-III tubulin and MBP in differentiated NPCs, which were consistent with the immunostaining results. However, the expression of GFAP in the Chase ABC-treated group showed no obvious change.