Anti at100

Twenty micrograms of protein of each sample were run on SDS-polyacrylamide gels (TGX gels, Bio-Rad), and transferred to nitrocellulose membranes by using the Trans-Blot® Turbo^™ Transfer System (Bio-Rad). The membranes were blocked with 5% BSA in Tris-buffered saline (TBS) plus 0.1% Tween-20 for 1 h, and then incubated with primary antibodies at 4°C overnight. The primary antibodies used were: anti-AT8 (Thermo Fisher, MN1020), anti-AT100 (Thermo Fisher, MN1060), anti-AT180 (Thermo Fisher, MN1040) and total Tau (C-terminal, Dako A0024). After washes with PBST, sections were incubated with Alexa 488- and 594-conjugated secondary antibodies (Invitrogen). The membranes were imaged using the ChemiDoc^™ system (Bio-Rad), and quantified with FIJI software.

All flies were age-matched for experiments. The whole-mount preparation of fly eyes and brains was performed as previously described^{46 (link)}.The following primary antibodies were used with the indicated dilutions: anti-lamin B1 (Sigma, 1:20), anti-GABA (GeneTex, 1:200), anti-human pan-tau (Dako, 1:200), anti-tau-C3 (Invitrogen, 1:200), anti-GFP (Abcam, 1:100), anti-AT8 (Thermo, 1:200), anti-AT100 (Thermo, 1:100), and anti-PHF1 (Abcam, 1:100). Alexa Fluor 488, Cy3, and Cy5 conjugated secondary antibodies (Jackson ImmunoResearch Laboratories) were used at 1:100 dilutions. F-Actin enriched rhabdomere and spots of aberrant actin accumulations were labeled by rhodamine-conjugated phalloidin (Sigma, 1:20) and Alexa Fluor 633-conjugated phalloidin (Thermo, 1:50), respectively. Samples were analyzed on Zeiss LSM510 or LSM800 confocal microscopes.