Fig. 1
From: Tau seeding activity in various regions of down syndrome brain assessed by two novel assays
Tau capture assay assessed the seeding activity of AD O-tau. a Tau level in AD O-tau was measured by immuno-dot blot. Various amounts of AD O-tau and tau39 (2N3R tau) were applied onto nitrocellulose (NC) membrane and developed with pan-tau antibody R134d. b–d. Tau151-391 was effectively captured by AD O-tau. Various amounts of AD O-tau were applied onto NC membrane and incubated with HEK-293FT cell extracts containing full-length (tau1-441) or truncated (tau151-391) tau40 (2N4R tau) tagged with HA. Captured HA-tau was developed with anti-HA followed with HRP-anti-mouse IgG (b). The level of tau in the cell lysates was analyzed by immuno-dot blots developed with anti-HA (c). The mean levels of captured tau1-441 or tau151-391 were plotted against the relative levels of AD O-tau (d). A.U., arbitrary unit. e AD O-tau could not recruit TDP-43 from cell lysate. NC membranes pre-applied with AD O-tau were incubated with 3R-tau151-391 or 4R-tau151-391 and TDP-431–414, followed with anti-HA and HRP-anti-mouse IgG. f Guanidine hydrochloride (GuHCl) killed the seeding activity of AD O-tau. NC membranes pre-applied with AD O-tau were treated with TBS, 6 N GuHCl, or 8 M urea for 2 h at RT. After washing with TBS, the membranes were subjected to tau capture assay or immuno-dot blots developed with R134d (pan-tau), PHF-1 (phospho-tau at Ser396/404), and TOMA1 (oligomeric tau)