Sample Preparation. With 3 mice in each group, the analytes were fixed in paraformaldehyde after cardiac perfusion. 30% sucrose was then added after 24 hours, and brain tissues were frozen sectioned with OTC embedded when sunk into the bottom of the bottle.
Proteinase K was added at 37°C after the brain tissues were washed three times with PBS with 5 mins each. 30 min later, LRP1 (USA, Abcam, ab28320, 1 : 70) and Aβ1–42 (USA, Abcam, ab10148, 1 : 100) were added equivalently after washing for three times with PBS as above. With incubation overnight at 4°C, the second antibodies (FITC (USA, Abcam, ab6785, 1 : 250) and Alexa Fluor® 647 (USA, Abcam, ab150079, 1 : 150)) were added after washing with the same method. With incubation for 1.5 hours at indoor temperature, DAPI (Beijing Zhongshan Golden Bridge, ZLI-9557) was added to fix the sections after washing as before. A laser scanning confocal microscope (FV1000, Olympus Corporation, Japan) was used for sections observation.