Immunoblot was performed as previously reported [26 (link)], with slight modification. Precast 4–20% tis-glycine or 10–20% tris-tricine gels were used (Bio-Rad). The iBlot™ Gel transfer device (Invitrogen) was used for transfer. The membranes were incubated in blocking buffer (Blocking Buffer for Fluorescent Western Blotting Rockland MB-070) for 1 h at room temperature. The following primary antibodies were used: 6E10 (Covance SIG-39300, 1:1000), APP, C-terminal (Sigma A8717, 1:1000), actin (Sigma A2066, 1:1000), actin (Sigma A3853, 1:1000), phospho-tau Ser404 (Thermo Fisher Scientific #44-758G, 1:2000), phospho-tau pThr181 (Thermo Fisher Scientific #701530, 1:1000), tau (Thermo Fisher Scientific #AHB0042, 1:2000). Antibodies were diluted in the blocking buffer, and membranes were incubated overnight at 4°C. The membranes were washed three times with TBST and secondary antibodies (Odyssey IRDye 680 or 800, all 1:10000) were applied for 1 h at room temperature. After washing four times, proteins were visualized using a Licor Odyssey Infrared imaging system. Blots were analyzed using ImageJ 1.47v software (National Institutes of Health). For the detection of Aβ, the membranes were microwaved in PBS to unmask the epitopes and increase the immunoblot sensitivity before blocking [69 (link)].