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L-soluble and sarkosyl-insoluble tau probed with antibodies against total tau (DAKO). Bar charts show sarkosyl-soluble and sarkosyl-insoluble tau as a proportion of tau in low speed supernatants as a measure of total tau. c Postmortem brain sections immunostained with an anti-tau (AT8) antibody show Braak staging of AD brain. NFTs are absent from age-matched control brain. CTRL: control (n = 5), Braak II AD (n = 4), Braak III AD (n = 3), Braak IV AD (n = 4), Braak V AD (n = 3), Braak VI AD (n = 5). Data is mean ?SEM. *p PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/17139194 regions of AD brain [35]. Following normalization of tau amounts to NSE, we found an increase in total tau protein in mid-late stage AD. Tau protein amounts were significantly increased in Braak stage IV and V tissues compared to control (p Capecitabine (P2), all of which were immunoblotted with antibodies against total tau and pSer396/404 (PHF1). These findings confirmed an increase in insoluble tau as a proportion of total tau in Braak stage V and VI tissues relative to earlier Braak stages and controls (Fig. 1b). Thus, the increase in total tau protein observed in Fig. 1a likely reflects the accumulation of this insoluble tau in tissue lysates, particularly since no changes in total tau mRNA have been reported in sporadic AD cortex [7, 28]. Immunohistochemical studies of fixed postmortem cortex labelled with the AT8 phospho-antibody are shown to confirm the Braak staging of these samples;Kurbatskaya et al. Acta Neuropathologica Communications (2016) 4:Page 5 ofthese show progressive appearance of characteristic tangle-like structures in Braak IV-VI tissues (Fig. 1c).Total APP amounts are increased in Braak stage II-III brainAmyloid precursor protein (APP) is a type 1 transmembrane glycoprotein that, in AD, is pathologically cleaved to give rise to A peptides of varying length [13]. We assessed amounts of APP holoprotein in postmortem cortex by probing blots with an antibody specific for Cterminal APP (6E10), which yielded two main bands at 106 and 113 kDa and a faint band at 130 kDa in late-stage AD brain, together characteristic of the three major APP isoforms found in human brain [14, 54], Fig. 2a). When standardized to NSE, total APP amounts were significantly increased in Braak stage II and III (p < 0.005) tissue compared to control, before returning to approximately control amounts in late (Braak IV-VI) stage AD (Fig. 2b). This finding extends previous studies which have sh.

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