Tau inclusions are not immunoreactive for ubiquilin 2.

<p>Double immunostaining for ubiquilin 2 and AT8 in an AD case (A), a G272V MAPT mutation carrier (B) and a sporadic PiD case (C). Detailed images of double labeling show neurons with AT8-positive tau inclusions (D-O). Images were spectrally unmixed, and shown separately and merged in artificial colors (ubiquilin 2: green; AT8: red). AD, Alzheimer’s disease; CA, cornu ammonis FTD, frontotemporal dementia; MAPT, microtubule-associated protein tau, Ub2, ubiquilin 2.</p

Ubiquilin 2 immunoreactivity in tauopathies.

<p>Representative immunohistochemical staining of ubiquilin 2 in the CA1 region of the hippocampus in a control (A) and an AD case (B). Detailed images showing plaque-like structures (C) and perisomatic granules (D) in an AD case. Ubiquilin 2 immunoreactivity in the subiculum of a sporadic PiD case (E), a G272V MAPT mutation carrier (F), and a PSP case (G). AD, Alzheimer’s disease; CA, cornu ammonis, FTD, frontotemporal dementia; MAPT, microtubule associated protein tau.</p

Additional file 2: Figure S2. of Activation of the unfolded protein response and granulovacuolar degeneration are not common features of human prion pathology

Immunohistochemical detection of PrP, pIRE1α, pPERK, and CK1δ in brain tissue of various human prion disease subtypes. Representative pictures of the immunohistochemical detection of PrP (3F4 antibody), pIRE1α, pPERK and CK1δ in frontal cortex sections of human prion disease patients with different disease subtypes, namely GSS (case #10), VPSPr (case #56), sCJD (case #27), panencephalopatic CJD (case #55), PrP-CAA (case #15), FFI (case #18), vCJD (case #21) and iCJD (#24), showing the absence of these UPR activation and GVD markers in the presence of PrP deposition. Immunohistochemical detection is visualized by DAB (brown staining) and nuclei are counterstained with haematoxylin (blue staining). Bar 200 μm. (PDF 2131 kb

QC expression is not upregulated by Aβ or the UPR.

<p>A. Differentiated SK-N-SH cells were treated with 1 µM and 2 µM oligomeric (O) and fibrillary (F) Aβ_1–42 for 24 h. Shown is the average + SD of normalized QC mRNA levels of triplicates from a representative experiment B. Differentiated SK-N-SH cells were treated with different UPR inducers, TM (0.2 µg/µl and 0.5 µg/µl), TG (1 µM and 2 µM) and DD (20 mM) for 16 h. Shown is the average + SD of normalized QC mRNA levels of n = 9 from 3 independent experiments. Normalized BiP mRNA levels are shown as positive control for UPR induction (n = 6 from two independent experiments). The expression levels were normalized to eEF2α mRNA, the levels in untreated cells are set to 1. Asterisks indicate a significant difference compared with control (*p≤0.01, **p≤0.001, ***p≤0.0001).</p

Post-mortem brain material used in this study.

<p>Listed are clinical diagnosis (CON = control, AD = Alzheimer’s disease), Braak stage, gender, age (in years), ApoE genotype, post-mortem interval (PMI, in hours: minutes).</p

Disturbed Ca²⁺ homeostasis increases QC expression and enzyme activity.

<p>A. Differentiated SK-N-SH cells were treated with different concentrations TG as indicated. Shown are the average + SD of normalized QC mRNA levels of n = 6 from two independent experiments. The expression levels were normalized to eEF2α mRNA, the expression levels in untreated cells are set to 1. B. Differentiated SK-N-SH cells were treated with 1 µM TG for 16 h. The enzymatic activity of QC activity was determined in protein lysates as described in materials and methods, activity in untreated cells is set to 1. Shown is average +SEM of n = 15 from 5 independent experiments. Asterisks indicate a significant difference compared to control (*p≤0.001, **p≤0.0001).</p

Primers and probes used for qPCR 480 light cycler.

<p>Probe numbers refer to numbers in the Roche universal probe library.</p

Additional file 3: Figure S3. of Activation of the unfolded protein response and granulovacuolar degeneration are not common features of human prion pathology

Immunohistochemical detection of pIRE1α and pPERK in AD and CJD hippocampus. Immunohistochemical detection for pIRE1α and pPERK was performed on an AD (case #57) and CJD (case #42) brain tissue with identical post-mortem handling and delay. Pictures from the subiculum are shown. Immunohistochemical detection is visualized by DAB (brown staining) and nuclei are counterstained with haematoxylin (blue staining). Bar 50 μm. (TIF 21347 kb

Additional file 1: Figure S1. of Activation of the unfolded protein response and granulovacuolar degeneration are not common features of human prion pathology

Semi-quantitative analysis of the Aβ (IC16) and phosphorylated tau (AT8) burden. Representative examples from our cohort showing the amount of Aβ deposits, small phosphorylated tau positive extracellular inclusions and neuritic changes related to prion disease and phosphorylated tau positive tangle-like structures, most often resulting from AD pathology, corresponding to the semi-quantitative scores +, ++ and +++. Aβ images were taken with the 5× objective, tau (positive inclusions/neuritic changes) images with the 10× objective, tau (tangle-like changes) images with the 20× objective. In Table 3 scores for each case are listed. Scores with a/, e.g. +/++, have a burden in-between the classes represented here. Brown staining with DAB, blue staining of the nucleus with haematoxylin. (PDF 1437 kb

Induction of c-fos by ER Ca²⁺ depletion precedes QC expression.

<p>Differentiated SK-N-SH cells were treated with 1 µM TG for 5 h and 16 h with or without pre-incubation with 5 µM BAPTA-AM (B) for 1 h. Shown are the average + SD of triplicates of normalized QC (A) and c-fos (B) mRNA levels from a representative experiment of three. The expression levels were normalized to eEF2α mRNA, the expression levels in untreated cells are set to 1. Asterisks indicate a significant difference compared to control (*p<0.05, **p≤0.02 ***p≤0.001).</p