Culture and transfection of HEK293T cells
By 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
Immunohistochemistry
By onThis protocol describes immunohistochemical staining of fixed brain sections.
Midbrain organoid differentiation in spinner flasks
By onMidbrain differentiation protocol using spinner flasks
Rab8a expression and purification
By onRecombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs
Single cell analysis of iPSC-derived midbrain organoids
By onThe following script was used for analysis of gene corrected (GC) versus GBA1 mutant (MUT) midbrain organoids. The purpose was to combine, filter, integrate, and identify clusters and differentially expressed genes sets. This is part of a Collection of protocols (dx.doi.org/10.17504/protocols.io.8epv593dng1b/v1) for the paper "Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function" (https://doi.org/10.21203/rs.3.rs-1521848/v1)
Immunohistochemistry using paraffin embedded tissue
By onImmunohistochemistry using paraffin embedded tissue
qPCR of α-synuclein, TNF and NF-κβ
By onA procedure for quantitative real time reverse transcription PCR of α-synuclein, TNF, and NF-κβ.
Subcellular and regional localization of mRNA translation in midbrain dopamine neurons
By onUsing, highly sensitive ribosome-bound RNA sequencing and imaging to characterize the translatome, the authors uncovered local mRNA translation of dopamine synthesis, release, and reuptake machinery in dendrites, but not axons.
Evaluation of pUb kinetics using 3D-SIM
By onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Assaying starvation-induced autophagy in HeLa cells
By onA method for assaying starvation-induced autophagy in HeLa cells that have been transfected with Halo-tagged constructs.
pCAG-ATG13 (363-517)-ULK1 (836-1050)-TSF
By onPlasmid for mammalian expression of ATG13 C-terminal and ULK1 MIT domain fusion complex.
Unconventional Initiation of PINK1/Parkin Mitophagy by Optineurin
By onThis work reveals that OPTN mitophagy initiation is mechanistically distinct and highlights the mechanistic plasticity of selective autophagy pathways.
Tet-off Full Length Tau (0N4R, P301L/V337M)
By onPlasmid: Tet-off Full Length Tau (0N4R, P301L/V337M). Associated with the following preprint: Saha et al., 2022 (published on biorxiv on February 19, 2022).
Live imaging and analysis to investigate phase separation properties of NEMO during mitophagy
By onProtocol for live imaging and analysis to investigate phase separation properties of NEMO during mitophagy.
Immunocytochemistry of cultured human Medium Spiny Neurons (MSNs)
By onThis protocol describes the immunolabelling of one or several protein targets on PFA-fixed cell culture on glass coverslips.
Accelerated Ovarian Failure AOF mouse model: a model of transitional menopause
By onThe novel model of AOF successfully replicates the human peri- and post-menopause stages, including erratic estrous cycles and hormone fluctuations, followed by undetectable estrogen levels.
