FIG. 6.

Effect of early DA treatment on differentiating H9 hESCs. (A) Time course of the experiment. Cells were treated with continuous DA during Stages 2–3 (days 1–11) of differentiation. (B, C) Phase-contrast images of colonies midway through the differentiation protocol (day 17, Stage 4). (B) Colony lacking neuroepithelial morphology. (C) Colony displaying primitive neuroepithelial morphology. Arrows indicate characteristic ridges between groups of elongated cells not seen in (B). (D) Quantitation of morphology midway through the differentiation protocol (day 17, Stage 4), % of colonies with neuroepithelial morphology were determined for each coverslip (n=8 slips per group). (E) Quantitation of the number of TH⁺ and TUJ1⁺ clusters per coverslip with early DA treatment. Cells were fixed on day 25 after 7 days in terminal differentiation media. Immunofluorescent staining for TH and TUJ1 was performed. Clusters of 10 or more TH⁺ cells and 20 or more TUJ1⁺ cells were counted. 7–8 slips per group. *Tukey test, P<0.05. **Tukey test, P<0.001. ***t-test P=0.0002. Error bars=SEM.