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Supplementary Figure 2

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posted on 27.11.2021, 19:12 by Jessica BinderJessica Binder

Supplementary Figure 2. Schematic showing the study design and overview of the data for the validation of predicted genes. A three-prong validation pipeline designed to validate the association of predicted genes with AD-related tau phenotype. The first set of validations investigate the mRNA levels of genes of interest (GOI; all twenty predicted genes) in human inducible pluripotent stem cell derived neurons (iPSNs) from sporadic AD (sAD2.1 – from Coriell Institute; # GM24666; 83 years old male) and control (AX0018 – from Axol Bioscience; #ax0018-kit; 74 years old male). The second set of validations involve mRNA and proteins analysis of each GOI from clinically diagnosed post-mortem brain samples (temporal cortices) from sporadic AD and non-demented healthy controls. The third set of validations include investigation of inflammation-induced tau phosphorylation following siRNA-mediated knockdown of GOI in SH-SY5Y human neuroblastoma cells. In this model, first the GOI is knocked down by siRNA followed by treating them with conditioned media (CM) derived from LPS (100 ng/ml)-activated microglia. Levels of phospho(p)-Ser199/pS202 (AT8 site) and pT231 (AT180 site) are assessed in SH-SY5Y cells.

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