Fig. 2. Treatment with KP or I3M increases neuronal differentiation of VM precursors, particularly into DA neurons. (A,B) Immunostaining with antibodies against nestin/Hoechst 33258 (A) and vimentin/Hoechst 33258 (B), showed a reduction in the number of neuronal precursors (nestin⁺) after treatment with I3M, with no effect on glial precursors (vimentin⁺). (C-F) 15 µM KP or 5 µM I3M increased the number of immature Tuj1⁺ (C,F) and mature MAP2⁺ (D,F) neurons and the proportion of TH⁺ DA neurons (E,F) after 3 days in vitro. Statistical analysis was performed using one-way ANOVA with Boneferroni's multiple comparison test (A,B) and two-tailed unpaired t-tests (C-E). Significance levels as for Fig. 1. Bar, 25 µm.