Figure 1

Relationship between PKCδ proteolytic activation and cell death during initial stages of H ₂ O ₂ -induced oxidative stress in N27 dopaminergic neuronal cell model. N27 dopaminergic cells were treated with H₂O₂ (100 μM) for 0-180 min and assayed for ROS generation using DCFDA dye (A) and for cytotoxicity using Sytox green dye (B). Non-linear regression graph from two or more independent experiments (n = 6-8). PKCδ cleavage was monitored by Western blot in a time dependent manner for 0 - 90 min in N27 dopaminergic cells treated with H₂O₂ (100 μM) (C). N27 dopaminergic cells were treated with H₂O₂ (100 μM) for 0, 30 or 60 minutes and PKCδ kinase activity was measured using [³²P] kinase assay; the bands were quantified for the graph (D). Data quantified from B and C were used to generate a bar graph and were compared with cytotoxicity and PKCδ cleavage following H₂O₂ exposure for 60 min. **, p < 0.01 as indicated by two-way ANOVA analysis using Bonferroni post test (E). N27 cells were transfected with 1 μM PKCδ siRNA and non-specific siRNA for 24 h and treated with 100 μM H₂O₂ and monitored for cytotoxicity using Sytox green dye, which showed significant protection from oxidative stress. ***, p < 0.001 denotes significant difference between non-specific siRNA- H₂O₂ and PKCδ siRNA H₂O₂-treated groups from two or more independent experiments (n = 6-8). Statistics were performed by one-way ANOVA analysis using Bonferroni post test (F).