FIGURE 5.

Juglone inhibits MPP⁺-induced expression of Pin1 in primary neuronal culture. Mesencephalic tissues from E15 mouse embryos were cultured and grown on laminin-coated coverslips. The neuronal cultures were treated with 10 μm MPP⁺ for 24 h in the presence or absence of 3 μm juglone. After culture, primary neurons were fixed with 4% paraformaldehyde and incubated with anti-TH and anti-Pin1 antibodies and viewed under a NIKON TE2000 fluorescence microscope. A, double labeling of TH and Pin1 in primary mesencephalic culture from substantia nigra is shown. M, MPP⁺. B, mesencephalic neuronal process lengths were quantified using MetaMorph image analysis software as mentioned “under Experimental Procedures.” C, shown is dopamine uptake assay from primary striatal culture. Results are the mean ± S.E. of at least three independent experiments. ***p < 0.001 versus control; ###, p < 0.001 versus MPP⁺; **, p < 0.01 versus MPP⁺.