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Fig. 3. GSK-3ß inhibition does not regulate cell survival or proliferation of VM precursors or MN9D cells but induces their morphological differentiation. (A) Treatment with 15 µM KP or 5 µM I3M did not change the total number of VM cells after 3 days in culture, as assessed by Hoechst 33258 staining. (B) Proliferation, as assessed by BrdU incorporation, was not affected by 15 µM KP, but was significantly decreased with 5 µM I3M. (C,D) Immunostaining with -Active Caspase 3 showed a decreased number of positive cells 3 days after treatment with 5 µM I3M, but not with KP. (E,F) Treatment of MN9D cells with 15 µM KP for 4 days did not alter the total number of cells (Hoechst 33258) or their proliferation (BrdU incorporation). (G) Treatment with 15 µM KP for 4 days induced morphological differentiation of MN9D cells. Statistical analysis was performed using two-tailed unpaired t-test (A, E and F), Kruskal-Wallis test (B) and one-way ANOVA with Boneferroni's multiple comparison test (C). Significance levels as for Fig. 1. Bar, 50 µm.

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