Tracking p14-induced cellular fusion through the visualization of cellular membranes and nuclei [graphic]

Milan Cuthbert
2017
Introduction Our lab has developed an alternative drug delivery platform that delivers cargo to cells of interest by causing fusion between the target cell and the drug delivery platform. This fusion bypasses the requirement of the drug to pass through the cell membrane. Our aim was to track cellular fusion by labeling the cell membrane and nuclei, inducing fusion, and then tracking the fusion over time. We also set out to create a movie showing said fusion. Methods QM5 cells were used in the
more » ... itial fusion functionality assay to track the effectiveness of p14 plasmid DNA to cause fusion. This provided the information for which DNA concentrations to use when tracking fusion in fluorescent HT1080 cells. Three fluorescent labels were used: PALM-GFP and PALM-tdTomato (membrane labels), and Hoechst 33342, a blue DNA stain. The membrane stains remained on the cells throughout the experiment. Plasmids were transfected onto cells and ~2hs later images were taken for 15 minute intervals. Acknowledgements:
doi:10.7939/r3zk5615n fatcat:7gxwip6ouzdcxj2dbmyi7hkhcm