FIG. 9.

Midbrain neuron markers upon early or late treatment with dopaminergic drugs. (A) Time course of the differentiation protocol showing treatment with DA, D1 antagonist SKF83566 [0.5 μM] (SKF8), D2 antagonist L741 [0.5 μM]. mRNA was extracted at 36 days (quantification), except for panel (D), where extraction was done on day 8. (B) rtPCR of PAX6 in human fetal brain tissue samples (CP, SVZ, Cx, and GE) and adult brain (hBR). (C) rtPCR of PAX6, 3 midbrain markers, and housekeeping gene ACTB during each differentiation stage. (D) qPCR of PAX6 in neuroepithelial colonies (control n=14; SKF83566 n=7). (E) qPCR of housekeeping gene (ACTB), neuronal markers (VGLUT1, GAD1, and βIII-Tub), and midbrain markers (GIRK2, LMX1A, and MSX1), in early and late treated cells. PCRs for VGLUT1 and MSX1 late treatment were repeated 4 times and the results averaged. Asterisks indicate significant reduction. Post hoc Tukey tests (SKF83566 P<0.01) and (L741 P<0.05). In panel (E₇) one-way ANOVA (P<0.05), but post hoc Tukey tests did not reveal any differences to the control group (for columns with error bars, n=4).