The Prion Protein N1 and N2 Cleavage Fragments Bind to Phosphatidylserine and Phosphatidic Acid; Relevance to Stress-Protection Responses
Fig 4
Serum deprivation causes changes to PS and PA in CF10 cells.
A. Laurdan GP changes following serum deprivation for 30 minutes as compared with benzyl alcohol (BA) and filipin III controls. n = 3. B. Live cell imaging of NBD-PS labelled CF10 cells. Image intensity is thresholds have been selected to view detail in the staining pattern and do not represent a comparison of fluorescence intensity. Scale bars = 20 μm. C. Fluorescence emission spectra of NBD-PS labelled cells following transfer into serum-free medium, scans were taken immediately after media replacement. D. Anisotropy of NBD-PS in CF10 cells with and without serum present. n = 3. E. Counts from magnetic separation of cells that have lost membrane asymmetry allowing them to bind PS at 5 and 15 minutes post serum withdrawal. n = 4. F. ROS production detected by DCF fluorescence when cells are serum-starved and with exposure to butan-1-ol to inhibit PLD activity. n = 4. G. Measurement of cellular phosphatidic acid concentration 30 minutes after commencing serum deprivation. n = 3. H. Measurement of phospholipase-D activity following 15 minutes serum starvation. n = 3. For all panels, *p<0.05, ***p<0.001.