Figure 2

PKD1 is highly expressed in dopaminergic neurons and activated during initial stages of H ₂ O ₂ -induced oxidative stress. Immunofluorescence analysis of N27 dopaminergic cells stained for native PKD1 using fluorescence and confocal microscopy. Nuclei were stained with Hoechst dye (A). Primary dopaminergic neurons staining for tyrosine hydroxylase (TH) obtained from the mouse substantia nigral region show co-localization of native PKD1 with TH. TH - Green, PKD1 native - Red, Nucleus - Blue, Yellow -Merge. Nuclei were stained with Hoechst dye (B). N27 dopaminergic neuronal cells were treated with or without H₂O₂ (100 μM) for 30, 60 or 90 min and probed for PKD1 activation loop phosphorylation pS744/pS748 and native PKD1 expression (C). N27 cells were treated with or without H₂O₂ (100 μM) and PKD1 kinase activity was measured by [³²P] kinase assay using syntide 2 substrate at 60 min (D). *, p < 0.05 denotes significant difference between untreated and H₂O₂ -treated groups.