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Fig. 1. Indirubin-3-monoxime (I3M) and kenpaullone (KP) increase the number of tyrosine hydroxylase positive (TH⁺) neurons in ventral mesencephalon (VM) precursor cultures. (A) Dose-response experiments in E14.5 VM precursors indicate that 15 µM KP and 3-5 µM I3M are optimal doses to increase TH⁺ cell numbers. Statistical analysis was performed using one-way ANOVA with Fisher's post-hoc test. ^*P<0.05; ^**0.01<P<0.001; ^***P<0.001. (B) Treatment of E14.5 VM precursors with 15 µM KP or 5 µM I3M for 24 hours decreases GSK-3ß protein levels as assessed by immunoblotting. ß-actin was used as a loading control. (C) TH/Hoechst 33258 immunostaining shows an increase in the number of TH immunoreactive cells 3 days after treatment with 15 µM KP or 5 µM I3M. Bar, 50 µm.

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