Published on 12th August 2016.
Wild type cells in the W303 strain background and W303 cells transformed with the relevant plasmids. Cells were grown in Yeast Nitrogen Base medium (1x Difco™ YNB base, 1x Formedium™ Complete Supplement Mixture, 5.0 g/L ammonium sulfate, 20 g/L glucose) to exponential phase. Plasmids used were YEplac195 and YEplac195-fps1∆1 for experiments with constitutively open Fps122. For all sampling cells were grown to mid-log phase in YNB medium. After addition of stress agent to cultures samples were taken at indicated time points with zero samples taken before addition of stressor. For hyper-osmotic stress sorbitol was added to the medium to a final concentration of 0.8 M or 1.0 M sorbitol depending on the experiment, for initial model validation 0.4 M NaCl was also used for hyper-osmotic stress. For the calcofluor treatment 0.11 μM (100 μg/mL) was added to the medium.
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We thank Alejandro Colman-Lerner for helpful comments on the manuscript. S.R.T. was supported by the Ministry of Education of Saxony-Anhalt (Research Centre Dynamic Systems: Biosystems Engineering (XD3639HP/0306 and CDS, MW- 21LMS 5) and the International Max Planck Research School (IMPRS) Magdeburg for Advanced Methods in Process and Systems Engineering. J.S. is supported by German Ministry for Education and Science (BMBF) (FKN 0315779 and 0316188E to JS). Work in the Hohmann and Klipp labs was supported by the European Commission, the UNICELLSYS project, No. 201142.