Live-cell imaging for synaptic vesicle precursors in human iNeuron axons
By savannah onThe authors describe procedure and equipment used for live-imaging of synaptic vesicle precursors. This was performed using DIV21 human iPSC-derived excitatory glutamatergic neurons. Equipment and software used varied based on scheduled upgrades to microscopy equipment during the course of this study.
Membrane interaction and mechanism of LC3 lipidation machinery in autophagy – Raw GUV data
By savannah onRaw GUV data of fluorescent protein imaged on a Nikon A1 confocal microscope with a 63 × Plan 359 Apochromat 1.4 NA objective. Three biological replicates were performed for each experimental 360 condition. Identical laser power and gain settings were used during the course of all conditions.
Microscopy-based bead protein-protein interaction assay
By savannah onThis protocol describes how to perform microscopy-based bead protein-protein interaction assay with GST- or mCherry-tagged proteins as baits and fluorescently-tagged proteins as preys. The protocol requires to have purified proteins and allows to monitor protein-protein interaction in an equilibrium state. The fluorescent signal can be quantified.
pCAG-ATG13 (363-517)-ULK1 (836-1050)-TSF
By savannah onMammalian expression of ATG13 C-terminal and ULK1 MIT domain fusion complex.
H9 ES AAVS-NGN2; FBXO7-/-
By savannah onH9 ES cells with AAVS1-NGN2 in which FBXO7 has been deleted using CRISPR.