Journal: Molecular Neurodegeneration
Article Title: BACE1 regulates expression of Clusterin in astrocytes for enhancing clearance of β-amyloid peptides
doi: 10.1186/s13024-023-00611-w
Figure Lengend Snippet: Clusterin underlies astrocyte endocytosis of Aβ in vitro. A Western blot of Bace1 -null and WT-primary astrocytes pretreated with either 80 pmol of Clu siRNA or 80 pmol of control scrambled siRNA and incubated with 2 μM of Aβ 42 for indicated times. Images indicate major bands for Aβ 42 (human, oligomeric) and GAPDH. B Quantification of Western blot Aβ 42 band intensity normalized to actin ( N = 3, * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001; One-way ANOVA with Tukey post testing comparing between samples. C Confocal imaging of astrocyte primary cultures from BACE1-null and WT and pretreated with either Clu or control scrambled siRNA then treated with 2 μM of Aβ 42 tagged with fluorescent Hilyte-555 for indicated incubation times. Stained with phalloidin (green) to mark F-actin and ToPro3 (blue) to mark nuclei. Scale bars represents 10 μm
Article Snippet: Knockdown of Clu in astrocyte cultures was attained by using Clusterin siRNA reagents (Santa Cruz Biotechnology, catalog no. sc-43689) or control scrambled siRNA (Santa Cruz Biotechnology, catalog no. sc-37007) according to manufacturer’s instructions.
Techniques: In Vitro, Western Blot, Control, Incubation, Imaging, Staining