Data for "Cellular control of viscosity counters changes in temperature and energy availability"
Raw data for "Cellular control of viscosity counters changes in temperature and energy availability". File contents are as follows: Raw Data 1.zip: Example FRAP movies Raw Data 2.zip: FRAP data for t-half Determination using easyFRAP Raw Data 3.zip: FRAP movies for diffusion coefficient determination using simFRAP Descriptions: Raw Data 1. Example FRAP movies, Related to Figures 2-5 and 7 and Figures S2-4, S6, and S7. Containing the following sub-folders: Movies 1. FRAP on GFP in cells transferred to multiple temperatures, non-steady state, Related to Figure 2B Movies 2. FRAP on GFP in cells in logarithmic growth at steady state for multiple temperatures, Related to Figure 2D Movies 3. FRAP on GFP in cells acutely heat shocked at 42°C in strains with and without impaired synthesis of glycogen, trehalose, or both, Related to Figure 3C Movies 4. FRAP on GFP in cells recovered from heat shocked with and without DAB and Validamycin A treatment, Related to Figure 3D Movies 5. FRAP on GFP in cells experiencing different conditions of nutrient deprivation, Related to Figures 4E and S4B Movies 6. In vitro FRAP on purified GFP in solutions of varying glycogen and trehalose content at 22°C and 40°C, Related to Figure 5A Movie 7. Magnetic beads puncture an inclusion in a lysate from cells heat shocked at 45°C for 30 minutes prior to lysis. Beads begin on one edge of the lysate and are drawn through the lysate by use of a strong magnet. and images are taken at 10x using brightfield illumination on an epifluorescent microscope, Related to Figure 6B-C Movies 8. FRAP on GFP in the Reg1 knockout strain in logarithmic growth and stationary phase, Related to Figures 6F and S6F. Movies 9. FRAP on GFP in wildtype yeast treated with the mitochondrial poisons sodium azide, sodium arsenite, or a combination of Antimycin A, Oligomycin A, and FCCP, with an untreated control, Related to Figure 7A. Raw Data 2. Data extracted from in vitro and in vivo FRAP movies for the determination of t-half values, Related to Main Figures 1-5 and 7 and Figures S2-4, S6, and S7. Data is in the form of excel spreadsheets of the format [column 1: time, column 2: intensity of bleached region, column 3: intensity of unbleached control region, and column 4: intensity of a non-fluorescent background region]. Spreadsheets were used as the input for the online tool easyFRAP (Koulouras et al., 2018). Related to Figures 1-7 and Figures S2-4,S6-7. Raw Data 3. Unprocessed FRAP movies used for determination of the diffusion coefficient of GFP in vitro and in vivo. Movies were used directly with the imagej software simFRAP (Blumenthal et al., 2015). Related to Figures 1 and 2.