Additional file 1 of Refining a mouse model of progressive supranuclear palsy through inoculation of human post-mortem brain-derived tau

Supplementary material 1: Figure 1: Schematic representation of PSP tau inoculation studies performed in 6htau mice. 1. The frontal cortex was obtained from a PSP postmortem brain. 2. Five different brain lysates were prepared. 3. Total tau was quantified in all lysates using an ELISA. 4. The yield of tau per gram of human brain was calculated for each of the lysates. 5. 6hTau mice were each inoculated in three subcortical nuclei implicated in early PSP. 6. Animal brains were examined for the presence of hyperphosphorylated tau (AT8 immunohistochemistry) and argentophilic neurofibrillary structures (modified Bielschowsy stain) at 3 and 6 months post inoculation. Figure 2: AT8 immunostaining in the frontal cortex of the PSP case used in the present study. AT8 immunostaining reveals abundant neuronal pathology (closed arrowhead) tufted astrocytes (open arrowhead) and oligodendroglial coiled bodies (dashed arrowhead). Scale bar 200um. Figure 3: Lack of 4R-tau seeding activity or AT8 immunopositivity in naive 6htau mouse brains at 3, 6 and 12 months of age. a) A 4R-Tau seeding amplification assay was performed in brain homogenates from a human PSP case, as well as 6hTau mice aged 3, 6 and 12 months of age. Immunostaining revealed a lack of AT8 immunopositivity throughout naïve 6hTau mouse brains at 3 (b), 6 (c) and 12 (d-f) months of age. Scale bar, show in f, represents 50um. Regions shown are caudate putamen (b-d), Globus pallidus (e) and substantia nigra pars compacta (f). Figure 4: AT8 immunostaining in 6hTau mice 6 months post inoculation with AD tau. Animals were inoculated in the caudate putamen (CPu), Substantia Nigra pars compacta (SNc) and Globus Pallidus (GP) with 1% SI tau from an AD case. Images show ipsilateral hemisphere (a-c), and both ipsilateral (right side) and contralateral hemispheres (left side) in d. Scale bar shown in d: 100um (a-c), 300um (d). Table shows semiquantitative Neuropathological scoring of AT8 immunopositivity. 0=none, 1=minimal, 2= mild, 3=moderate, 4=severe. * represents injection sites.