SNOC and H
have opposing effects on BACE1 enzymatic activity and subsequent Aβ generation. (A and B) Effect of nitrosative and oxidative stresses on BACE1 activity was measured by a fluorescent based β-secretase activity assay kit (Sigma). Different doses of SNOC and H2O2 were added to N2a-695 stable cells expressing human APP695 proteins, for 1 hr. Subsequently, cells were lysed by lysis buffer for measurement of BACE1 activity. (C) For further investigation of the effect of SNOC and H2O2 on Aβ generation, conditioned media was collected after 4 hrs incubation with OPTI-MEM media from cells treated with various concentrations of SNOC and H2O2 for 1 hr. Then, secreted Aβ peptides were immunoprecipitated with 6E10 antibody and protein G/A beads to measure secreted Aβ peptides.