Culture and transfection of HEK293T cells
By savannah onThis protocol describes a standard procedure culturing and transfecting HEK293T cells. Protocol overview: A. Culturing HEK293T cells; B. Transfection of HEK293T cells with Lipofectamine 2000.
CRISPResso analysis
By savannah onThis protocol describes the process of using CRISPResso to analyze Next Generation Sequencing (NGS) data from CRISPR-targeted, genome-edited samples.
Coating plates
By savannah onThis protocol describes the process of coating plates using either VTN, Matrigell, or Geltrex for use in culturing of feeder-free human pluripotent stem cells (hPSCs).
Amplifying the target genomic region by PCR
By savannah onThis protocol describes a procedure for amplifying targeted genomic regions using a PCR reaction.
Adapting hPSCs cultured on MEFs to feeder-free system
By savannah onThis protocol describes the procedure of adapting human pluripotent stem cells (hPSCs) to feeder-free culturing conditions using mTeSR-plus or StemFlex.
AAVS1 Knock-in
By savannah onThis protocol describes the standard procedure to knock-in constructs to the AAVS1 safe harbor locus in hPSCs.
Western blotting to detect ATP13A2 and ATP13A3
By savannah onProtocol to detect ATP13A2 and ATP13A3 via Western blotting.
Microscopy-based mitochondrial morphology measurements in iNeurons
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based mtDNA turnover measurements in HeLa and iNeurons
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based measurements of p62 recruitment in HeLa
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based pUb-coverage measurements of mitochondria in iNeurons
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based evaluation of Parkin translocation and mitophagy in FBXO7-/- cell lines
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Immunocytochemical analysis
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Evaluation of pUb kinetics using 3D-SIM
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Cell culture, transfection, and imaging
By savannah onThis protocol details the general preparation of cells for imaging and also for imaging experiments involving cellular hypotonic shock and cytosolic Ca2+ changes as they were performed in https://doi.org/10.1083/jcb.202010004.
Cell culture, transfection, and imaging
By savannah onThis protocol describes general procedures for culturing HeLa cells, transient transfection, and imaging using an Andor Dragonfly spinning disk confocal system.
Protocol collection for “PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons and HeLa cell systems”
By savannah onCollection of protocols associated with preprint (https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1.full). Protocols: Microscopy-based evaluation of Parkin translocation and mitophagy in FBXO7-/- cell lines Microscopy-based pUb-coverage measurements of mitochondria in iNeurons Microscopy-based measurements of p62 recruitment in HeLa Microscopy-based mtDNA turnover measurements in HeLa and iNeurons Microscopy-based mitochondrial morphology measurements in iNeurons
Flow cytometry-based measurement of mitophagic flux
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Evaluation of mtKeima foci in induced neurons (iNeurons)
By savannah onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
LC3-lipidation-assay
By savannah onProtocol for an in vitro LC3 lipidation assay using purified proteins and synthetic liposomes.
