Figure 2
Intracellular localization and pH calibration of sfpHluorin and ER-sfpHluorin. Fluorescence microscopy images of sfpHluorin (A) and ER-sfpHluorin (B) in S. cerevisiae cells. Scale bars correspond to 2 µm. pH calibration with permeabilized S. cerevisiae cells expressing sfpHluorin (C) and ER-sfpHluorin (D). The emission intensity at 512 nm with an excitation wavelength of 390 nm was divided through the emission intensity (512 nm) with an excitation wavelength of 470 nm yielding the ratio R390/470. Mean and standard deviation of biological triplicates are shown (mean ± s.d.). The graphical nonlinear fit (sigmoidal dose-response) was performed with the GraphPad Prism software. The best-fit values and the standard error of the fit can be found in the Supplementary Table S4A and B.
