Anti app c terminal

The fixation and staining of neuronal cultures were performed as described above. Primary antibody incubation was conducted with anti-DDX24 (1:100) HPA 002554 (Sigma-Aldrich, Saint Louis, MO, USA) and anti-APP C-terminal (1:200), C1/6.1 (BioLegend, San Diego, CA, USA) ON at 4 °C. Secondary antibodies used were Alexa Fluor 555 (1:1000), Alexa Fluor 647 (1:1000), and phalloidin (1:200) in 3% NGS in PBS.

Brain samples were homogenized in ice‐cold RIPA lysis buffer. Protein samples were loaded on a 4%–20% SDS‐polyacrylamide gel and then transferred onto nitrocellulose (NC) membranes. After blocking with 5% fat‐free milk, the NC membranes were incubated with the following primary antibodies overnight at 4°C: anti‐APP C‐terminal (1:1000, Biolegend); anti‐6E10 (1:400, Biolegend); anti‐RAGE (1:1000, Millipore); anti‐LRP‐1 (1:1000, Abcam); anti‐pS396‐tau (1:1000, Abcam); anti‐pT231‐tau (1:1000, Signalway); anti‐Tau5 (1:1000, Millipore); anti‐Synapsin‐1 (1:1000, Abcam); anti‐PSD95 (1:1000, Millipore); and anti‐β‐actin (1:2000, Origene). The membranes were incubated with the corresponding IRDye 800 CW‐conjugated secondary antibodies and scanned using an Odyssey fluorescent scanner. Relative band intensities were normalized to the band intensity of the internal reference protein for analysis.