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Reaction efficiency calculus was done using the amplification curve fluorescence, analyzing each tube separately as described by Liu and Saint (2002) [ 42].
We thank Dr. Hironobu Fujiyoshi and Dr. Shoichi Shimizu, Chubu University, for providing the fluorescence analyzing software FL ver. 1.0.0.1.
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For this, cells were plated on FITC-gelatin and disappearance of fluorescence analyzed.
Cells were processed as described above but fluorescence was analyzed using a fluorescence microplate reader.
Fluorescence was analyzed with a confocal fluorescence microscope (Zeiss LSM 510 Meta, Jena, Germany).
Cell fluorescence was analyzed at 488-nm excitation and applied to standard fluorescence compensation.
The relative intensity of 7-hydroxycoumarin fluorescence was analyzed.
GFP relative fluorescence was analyzed for every time point and compared to the control strain (w1.1).
Cells were harvested 2 days after transfection and the fluorescence was analyzed using flow cytometry.
Relative Nile Red fluorescence was analyzed using a BD FACSCanto II flow cytometer (BD Biosciences, San Jose, CA, USA).
GFP fluorescence was analyzed by flow cytometry.
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