Tau localization in hemi-synapses. (A) Immunofluorescence detection by DA9 (total tau), PHF1 (pS396/pS404 tau), and Alz-50 (misfolded tau) in isolated hemi-synapses, overlaid with brightfield (BF) images, from control and AD subjects. VGluT1/MAP2 are presynaptic/postsynaptic markers. (B) Quantification of tau immunofluorescence in scattered hemi-synapses. 51 hemi-synapses per subject; 4 human subjects per group, except for control PHF1 and control Alz-50 experiments which had only 3 subjects. Statistical results (n.s. not significant, *p < 0.05, **p < 0.01) are shown for two-way ANOVA (pre/post vs. Con/AD, black line; pre/post vs. PHF1/Alz-50, red line).