Additional file 1 of Tyrosine phosphatase STEP61 in human dementia and in animal models with amyloid and tau pathology
Creators: name: Taylor, Deonne
name: Kneynsberg, Andrew
name: van Roijen, Marloes
name: Götz, Jürgen
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Neuroscience
subject: 111714 Mental Health
subject: FOS: Health sciences

description: Additional file 1: Materials and methods. Methods. Reagents and antibodies, human samples, experimental mice, tissue processing, sequential protein fractionation, subcellular fractionation, western blots, statistical analysis; Materials. Human tissue samples. Table S1. Human patient information for clinically diagnosed FTD, AD, MCI and healthy control patients. Figure S1. Changes in STEP61 in the cortex of AD and FTD patients. A Quantification of levels of STEP61, its activity, PSD-95 and tau normalised to actin in AD (n=6), MCI (n=7) and healthy control (n=8). Fold change of the healthy control. One-way ANOVA post-hoc multiple comparisons correction performed with Tukey’s test, active STEP p=0.1093, total STEP p=0.0919, PSD-95 p=0.1144, tau p=0.8275. B Quantification of levels of STEP61, its activity, PSD-95 and tau normalised to actin in FTD (n=8) and healthy control (n=8) cortex, plotted by Braak staging, fold change of healthy controls. Analysed with unpaired Student’s t-test used for each quantification, active STEP p=0.9947, total STEP p=0.9098, PSD-95 p=0.9312, tau p=0.7568. Figure S2. No correlation detected between post mortem delay and STEP61 activity or expression. To determine whether post mortem delay effected the phosphorylation state of STEP61, as dephosphorylation results in STEP61 activation. Long delays can result in degradation of proteins or dephosphorylation by endogenous phosphatases, however we observed no correlation between post mortem delay and STEP61 activation or expression or PSD-95 expression. Colour coded to show clinical diagnosis of patients. A Correlation between active STEP61 expression and delay (hrs) of all patients. Correlation analysis by Pearson’s correlation coefficient, all grouped R2= 0.01425. B Correlation between total STEP61 expression and delay (hrs) of all patients. Correlation analysis by Pearson’s correlation coefficient, all grouped R2= 0.04894. C Correlation between total PSD-95 expression and delay (hrs) of all patients. Correlation analysis by Pearson’s correlation coefficient, all grouped R2= 0.09331. Figure S3. Correlation detected between PSD-95 expression and STEP61 expression in AD and FTD cohorts. Correlation between the expression of total STEP and PSD-95 expression in AD and FTD human cohorts. Shows decrease in STEP61 expression correlates with reduced PSD-95 expression. Colour coded to show clinical diagnosis of patients. A Correlation between total STEP61 expression and PSD-95 of all AD (blue), MCI (green) and control (black) patients. Correlation analysis by Pearson’s correlation coefficient, R2= 0.5297. B Correlation between total STEP61 expression and PSD-95 of all FTD-tau (red) and control (black) patients. Correlation analysis by Pearson’s correlation coefficient, R2= 0.3604. Figure S4. Fractionation protocol quality was identified by extrasynaptic enriched synaptophysin and synaptic enriched PSD-95. Immunoblot of the extrasynaptic and synaptic fractions of 6-month-old APP23 mice (n=4) and their WT littermate controls (n=4). Extrasynaptic fractions were enriched for synaptophysin, whereas the synaptic fraction was enriched with the post-synaptic marker PSD-95.