Hunter Medical Research Institute

4 ARTICLES PUBLISHED IN JoVE

Biology

Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell Lines

Brianna C. Morten^1,2, Rodney J. Scott^1,2,3, Kelly A. Avery-Kiejda^1,2

¹Medical Genetics, Hunter Medical Research Institute, ²Priority Research Centre for Cancer, School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, ³Pathology North, John Hunter Hospital

This protocol describes the use of three different methods for analyzing cell proliferation in breast cancer cell lines. This includes the use of conventional cell counting, luminescence-based cell viability, and cell counting through the use of a cell imager. Each offers advantages for the reproducible measurement of cell proliferation.

Developmental Biology

Analysis of Epididymal Protein Synthesis and Secretion

Wei Zhou^1,2, Petra Sipilä³, Geoffry N. De Iuliis^1,2, Matthew D. Dun^2,4, Brett Nixon^1,2

¹Priority Research Centre for Reproductive Science, School of Environmental and Life Sciences, University of Newcastle, ²Hunter Medical Research Institute, ³Department of Physiology, Turku Center for Disease Modeling, Institute of Biomedicine, University of Turku, ⁴School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle

Here, we report the immunofluorescence localization of dynamin to illustrate the protocols for the detection of proteins in paraffin-embedded mouse epididymal sections and those of an immortalized epididymal cell line (mECap18). We also describe the protocols for the isolation of secretory proteins from both epididymal fluid and conditioned cell media.

Cancer Research

A Simple Migration/Invasion Workflow Using an Automated Live-cell Imager

Xiajie Zhang^1,2, Brianna C. Morten^1,2, Rodney J. Scott^1,2,3, Kelly A. Avery-Kiejda^1,2

¹Medical Genetics, Hunter Medical Research Institute, ²Priority Research Centre for Cancer Research, Innovation and Translation, School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, ³Pathology North, John Hunter Hospital

The current protocol describes an integrated method investigating cancer cell migration and invasion on a single platform in real-time, providing an easily reproducible and time-efficient option to study cell mobility and morphology.

Biochemistry

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins

Wei Zhou¹, Hiroyuki Takeda²

¹Graduate School of Medicine, Ehime University, ²Proteo-Science Center, Ehime University

This protocol describes an efficient cell-free method for production of high-quality proteoliposome by bilayer-dialysis method using wheat cell-free system and liposomes. This method provides suitable means for functional analysis of membrane proteins, drug targets screening, and antibody development.