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Effects of PS2 Mutation on BACE1, BACE2, p-ERK Expression, and Oxidative Stress in PC12 Cells

This study investigates the impact of presenilin 2 (PS2) mutations on various cellular parameters in transfected PC12 cells. We analyzed BACE1 and BACE2 expression as well as phosphorylated ERK (p-ERK) levels using immunoblotting. Additionally, β-secretase activity, reactive oxygen species (ROS) generation, and lipid peroxidation products were assessed. Significant differences in these parameters were observed between cells expressing mutant PS2 and those expressing wild-type PS2 or a control vector. Results highlight the oxidative stress induced by PS2 mutations.

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Effects of PS2 Mutation on BACE1, BACE2, p-ERK Expression, and Oxidative Stress in PC12 Cells

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  1. Supplementary Fig. 2. Increase of BACE1, BASE2 and p-ERK expression, β-secretase activity, ROS generation and lipid peroxidation by PS2 mutation. A-D: Immunoblots of transfected PC12 cells; PC12/Neo, PC12/PS2wt and PC12/PS2mt, were probed with specific antibody for BACE1 and BACE2 (A) and p-ERK (C). β-actin protein was used as an internal control. β-secretase activity in the transfected PC12 cells was measured as described in Materials and Methods (B). D: Intracellular ROS levels were determined in transfected PC12 cells by measuring DCF fluorescence. E: Lipid peroxidation was determined by measuring the lipid peroxidation product; HNE-His adducts. Significant differences from neo or (*p<0.05), or PS2wt-Tg. Data were assessed by one way ANOVA followed by Dunnett’s post hoc test. Panel (A) is representative of 3 different experiments with duplicates. The data are mean ± SD of 3 experiments with duplicates (B-E). Significant differences from PC12 cells expressing vector alone (neo) (*p < 0.05) or PC12 expressing PS2wt (#p < 0.05) were assessed by one way ANOVA followed by Dunnett’s post hoc test.

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