Figure 7
Steady-state protein expression levels in mouse brain samples. Protein extracts were obtained from APP/PS1 and wild-type mice brain samples stored in TRIzol®-chloroform samples at −80°C for 3 months. (A) Sixty μg of total protein extracts from wild-type (WT) and APP/PS1 mice hippocampus were electrophoretically separated in 10-20% Tris-Tricine gels, transferred to nitrocellulose membrane and evaluated for the steady-state protein expression levels of human APP, APP-C-terminal fragment β (CTF-β) and amyloid-β (Aβ) peptide. Ponceau S staining was used as loading control. (B) Sixty μg of total protein extracts from WT and APP/PS1 mice hippocampus and frontal cortex were separated in 6 and 12% SDS-PAGE, transferred to nitrocellulose membrane and evaluated for the steady-state protein expression levels of apoE and SORLA. β-Actin and/or Ponceau S staining were used as loading controls. (C) Ten ng of total protein extracts from APP/PS1 mice hippocampus and frontal cortex (H and FC, respectively) were used to measure human total Aβ1–40 or Aβ1–42 content by sandwich ELISA, and to determine individual Aβ1–42 to Aβ1–40 ratio. Data are mean ± SEM from 5 mice in each brain region.