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2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Detection of In Situ Protein-protein Complexes at the Drosophila Larval Neuromuscular Junction Using Proximity Ligation Assay

Simon Wang¹, SooHyun Yoo¹, Hae-yoon Kim¹, Mannan Wang², Clare Zheng², Wade Parkhouse², Charles Krieger², Nicholas Harden¹

¹Department of Molecular Biology and Biochemistry, Simon Fraser University, ²Department of Biomedical Physiology and Kinesiology, Simon Fraser University

This protocol demonstrates how Proximity Ligation Assay can be used to detect in situ protein-protein interactions at the Drosophila larval neuromuscular junction. With this technique, Discs large and Hu-li tai shao are shown to form a complex at the postsynaptic region, an association previously identified through co-immunoprecipitation.

Medicine

Busulfan as a Myelosuppressive Agent for Generating Stable High-level Bone Marrow Chimerism in Mice

Kyle Peake¹, John Manning¹, Coral-Ann Lewis^1,2, Christine Barr¹, Fabio Rossi², Charles Krieger^1,3

¹Department of Biomedical Physiology and Kinesiology, Simon Fraser University, ²The Biomedical Research Centre, University of British Columbia, ³Division of Neurology, Department of Medicine, Neuromuscular Disease Unit, VHHSC

We describe a protocol whereby busulfan conditioning permits the bone marrow of a recipient mouse to be replaced with bone marrow cells from donor mice ubiquitously expressing green fluorescent protein, in the absence of irradiation. This technique is useful to study bone marrow cell accumulation in the central nervous system.