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Case Western Reserve University School of Medicine

4 ARTICLES PUBLISHED IN JoVE

Medicine

Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain

Xiangzhu Xiao¹, Jue Yuan¹, Wen-Quan Zou^1,2

¹Department of Pathology, National Prion Disease Pathology Surveillance Center, Case Western Reserve University School of Medicine, ²Department of Neurology, National Prion Disease Pathology Surveillance Center, Case Western Reserve University School of Medicine

A new species of cellular prion protein (PrP^C) has recently been identified in uninfected human brains using the methods described here. These methods can be used to isolate various PrP species, while some of them are also useful in isolating other misfolded protein aggregates from human brains.

Biology

Flow Cytometry-based Purification of S. cerevisiae Zygotes

Serendipity Zapanta Rinonos^1,2, Jeremy Saks¹, Jonida Toska³, Chun-Lun Ni^1,2, Alan M. Tartakoff^1,2

¹Department of Pathology, Case Western Reserve University School of Medicine, ²Cell Biology Program, Case Western Reserve University School of Medicine, ³Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine

To purify zygotes of S. cerevisiae, haploid cells of opposite mating type were engineered to express red or green fluorescent proteins, co-incubated to allow zygote formation, and fractionated using a flow cytometry-based protocol. The highly-enriched fraction enables subsequent "-omic" studies, recovery of initial progeny, and systematic investigation of zygote morphogenesis.

Biochemistry

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Ryan W. Clinton¹, Jason A. Mears¹

¹Department of Pharmacology, Center for Mitochondrial Diseases, The Cleveland Center for Membrane and Structural Biology, Case Western Reserve University School of Medicine

This paper describes a method for assessing the interactions and assemblies of integral membrane proteins in vitro with various partner factors in a lipid-proximal environment.

Cancer Research

Flow Cytometry-based Drug Screening System for the Identification of Small Molecules That Promote Cellular Differentiation of Glioblastoma Stem Cells

Raffaella Spina¹, Dillon M. Voss¹, Laura Asnaghi², Andrew Sloan^1,3, Eli E. Bar¹

¹Department of Neurological Surgery, Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, ²Department of Pathology, Johns Hopkins University, School of Medicine, ³Department of Neurological Surgery, University Hospital-Case Medical Center, Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine

An efficient screening protocol is presented for the identification of small molecules that promote astroglial differentiation in glioblastoma stem cells (GSCs). The assay is based on a stem cell differentiation reporter whereby the expression of the enhanced GFP (eGFP) is driven by the human GFAP promoter.