Generation of CRISPR constructs
By quincy.tichenor onThis protocol details the procedure of generation of CRISPR constructs.
Expression and purification protocol of ULK1 Complex wt or K46I mutant
By quincy.tichenor onThis protocol describes the expression and purification of ULK1 Complex wt or K46I mutant.
Determination of edits in CRISPR-edited cell lines by sequencing
By quincy.tichenor onThis protocol details the procedure of determination of edits in CRISPR-edited cell lines by sequencing.
Workflow for proteomic analysis of purified lysosomes in cells lacking GRN V.3
By quincy.tichenor onLyso-IP is a purification method of lysosomes, which can then be used for proteomic analysis or for metabolomic analysis. The Lyso-IP is adapted from a previous reported method. Here we also describe processing steps using proteomics after lysosome purification in the context of HeLa cells lacking the GRN gene.
Tomogram data of TauRD-Y forms amyloid-like aggregates in cells
By quincy.tichenor onTomograms: The tomograms shown in Fig. 1d and Fig. 3c are available in the EMDB https://www.ebi.ac.uk/emdb/ through the following information: EMD ID: EMD-13739 (Fig. 1d) and EMD-13740 (Fig. 3c) https://www.ebi.ac.uk/emdb/EMD-13739 https://www.ebi.ac.uk/emdb/EMD-13740 Associated with the following preprint: Saha et al., 2022 (published on biorxiv on Feb 19th, 2022) https://www.biorxiv.org/content/10.1101/2022.02.18.481043v1.full
Primary neuronal cultures
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Native-PAGE analysis of VCP hexamer
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Lentivirus production for primary neuron transduction
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Immunological detection of autophagy and mTORC1-related proteins
By quincy.tichenor onA general protocol for immunological detection by Western blotting MTOR, MTOR (pS2448), ULK1, ULK1 (pS757), p70S6K, p70S6K (pT389), SQSTM1, CALCOCO2, MAP1LC3B, GABARAP, TFEB, TFE3, PGRN, HSP90, and PCNA.
Immunohistochemistry of rTg4510 mouse brain
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Detection of Tau ubiquitination
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Biochemical detection of aggregated Tau
By quincy.tichenor onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Genome-wide determinants of mortality and clinical progression in Parkinson’s disease – Summary statistics (Dataset)
By quincy.tichenor onSummary statistics from "Genome-wide determinants of mortality and clinical progression in Parkinson’s disease". medRxiv: https://www.medrxiv.org/content/10.1101/2022.07.07.22277297v1
Code for making forest plots for top GWAS loci
By quincy.tichenor onThis repository contains the script used to create forest plots for top loci .
Membrane remodeling properties of the Parkinson’s disease protein LRRK2
By quincy.tichenor onPreprint: The authors examine how purified LRRK2 directly binds acidic lipid bilayers in a cell-free system and can deform them into narrow tubules in a guanylnucleotide-dependent but ATP-independent way.
Cellular senescence: a key therapeutic target in aging and diseases
By quincy.tichenor onReview: This review discusses the role of SnCs in aging and age-related diseases, strategies to target SnCs, approaches to discover and develop senotherapeutics, and preclinical and clinical advances of senolytics.
Disruption of lysosomal proteolysis in astrocytes facilitates midbrain proteostasis failure in an early-onset PD model
By quincy.tichenor onPublished: Accumulation of advanced glycation end products (AGEs) on biopolymers accompany cellular aging and drives poorly understood disease processes. Here, authors studied how AGEs contribute to development of early on-set Parkinson’s disease (PD) caused by loss-of-function of DJ1, a protein deglycase. View original preprint.
Mouse Stereotaxic Surgery
By Michelle onThis protocol describes the steps for performing stereotaxic surgery in mice. It is applicable to intracranial injections (e.g. virus, drug) and placement of implants (e.g. optical fibers, electrode arrays) into targeted regions of mouse brains.
Optogenetic Manipulation (Mouse)
By Michelle onThis protocol describes the steps for in vivo ontogenetic manipulation in mice, including assembly of fiber-ferrules, surgical implantation of fibers, and testing procedure.
