Relationship between PKCδ proteolytic activation and cell death during initial stages of H
-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).