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Novel Fluorescent Probes for the Oxidative Milieu

(cfSGFP2), 555 nm (cgfTagRFP) or 584 nm (cgfmKate2) was                                    (DOC)
plotted. The best-fits to Henderson-Hasselbalch equation were
determined as shown in Table.                                                              Acknowledgments
(TIF)
                                                                                           We thank all members of the Departments of Cell Science and Anatomy
Video S1 Live cell imaging of ss-cfSGFP2-prion fusion                                      and Histology of Fukushima Medical University. We also thank Dr.
protein. COS7 cells transiently expressing ss-cfSGFP2-prion was                            Yasuteru Urano at Tokyo University for a series of advice on measurement
exposed to 0.5 mM CuSO4 in phosphate-buffered saline at                                    of photochemical properties, and appreciate Hamamatsu Phononics for
pH 5.0 for 3 min at room temperature and images were                                       help in the measurement of quantum yield. We thank P. H. Cameron
continuously recorded at 32uC at 1 frame per min. The z-position                           (McGill University, Montreal, Canada) for her help in the preparation of
of images from an objective lens was set constant using the                                this manuscript.
PerfectFocus function of A1R confocal microscopy. Apparent
apoptotic cell death after internalization of large aggregates was                         Author Contributions
observed in some cells (left end). The width and height of images
were 81.9 mm.                                                                              Conceived and designed the experiments: IW. Performed the experiments:
(MP4)                                                                                      TS SA MT CS HE. Analyzed the data: IW TS HH KH. Contributed
                                                                                           reagents/materials/analysis tools: TS SA MT TH. Wrote the paper: IW
Table S1                                                                                   KH.

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PLoS ONE | www.plosone.org                                                                 15 May 2012 | Volume 7 | Issue 5 | e37551
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