Fig. 7

Using synaptoneurosomes to enrich and analyze synaptic proteins by western blotting. a Representative enrichment blot showing the exclusion of nuclear histone (17 kDa) from the synaptoneurosome preparation and retention of synapsin (40-80 kDa). b GluN2B western blot from the LBC1936 (black bars) and AD (grey bars) preparations. To assess protein integrity and control for post-mortem degradation [52], band2 (black arrow; 150 kDa) was divided by band1 (grey arrow; 170 kDa) to generate a ratio, and a value ≥1 (red dashed line) is achieved by all samples except the LBC1936 EC (asterix). c Representative synaptophysin (40 kDa) blot of LBC1936 synaptoneurosomes and AD samples. d Representative total tau (Tau13; 45–60 kDa) blot of LBC1936 samples and AD samples. e Phosphorylated-tau (PHF1; 50–65 kDa) blot shows almost exclusive expression in the AD samples compared to the LBC1936 synapses. f Representative VDAC (29–32 kDa) blot of LBC1936 and AD synaptoneurosomes. GAPDH (36 kDa) or β-actin (42 kDa) was run as a loading control. Histogram represents the mean of three experimental repeats and the bars represent the quantification of the bands directly above