Fig. 1

The scheme of the experimental workflow. Human AD brain-derived tau inocula were obtained from AD patients’ frontal cortex, tissue was homogenized, and tau was enriched from sarkosyl-insoluble fraction by sodium phosphotungstate. Tau concentrations and conformational properties in samples were evaluated by conformation-dependent immunoassay (CDI) and conformational stability assay (CSA). Mouse primary neurons were inoculated with human AD-tau samples of the same tau concentration at 7DIV for the maximum inoculation time point at 21DIV to investigate the template propagation of tau misfolding and aggregation. The viability and cytotoxicity were performed to assess tau concentration working range. By confocal microscopy, we investigated mouse tau aggregation after 14 days of inoculation, cell morphology, and the effects on synapse levels. Inoculated neuronal cultures were also lysed, and the concentration, aggregation rate, and conformational properties of sarkosyl-insoluble tau were measured from pellet fractions by CDI and western blots. The supernatants were used in western blot analysis of synaptic markers