Figure 5. Starvation induces a constriction in cell size and an expansion in vacuolar volume (Figure 5—figure supplements 1 and 2).

(A) Nuclear volume after acute glucose starvation. Histograms of nuclear volumes measured by reconstruction from three-dimensional image stacks using Imaris. The p-value resulting from a two-tailed t-test on the average volume in each condition (unpaired values assuming equal variance) is p<0.001. (B) Histograms of cell volumes of log-growing and acutely starved yeast cells. Log-growing cells, (+) glucose, were acutely starved of glucose, (–) glucose, and cell volume was measured using a Beckman Coulter Multisizer 3 (see 'Materials and methods'). Approximately 50,000 cells were measured for each condition. (C) Cytoplasmic free GFP (green) and vacuolar membrane protein Vph1-mCherry (magenta) fluorescence images. Scale bar: 10 μm. (D) Quantification of vacuole-to-cell volume ratio. Error bars represent standard errors about the mean (N = 3 independent experiments with > 55 cells per experiment). (E) Log-log MSD plot of the POA1 locus after treatment with increasing concentrations of NaCl. Cells were imaged approximately 10 min after hyperosmotic shock. (F) Log-log MSD plot of the GFA1 mRNP after treatment as described in (E).

DOI: http://dx.doi.org/10.7554/eLife.09376.014

Figure 5—figure supplement 1. Histograms of cell volumes after various treatments.

(A) Cells were treated with K+Sorbate as in Figure 4 and cell volume measured as in Figure 5B. (B) Cells were starved of glucose in either low or high pH (5.0 or 7.4) and cell volume was measured.

DOI: http://dx.doi.org/10.7554/eLife.09376.015

Figure 5—figure supplement 2. Hyperosmotic shock increasingly confines macromolecular mobility without affecting intracellular ATP concentrations or intracellular pH.

(A) Intracellular ATP concentrations after treatment with either 0.4 M or 0.8 M NaCl. Cells were treated as described in Figure 5E and intracellular ATP concentrations were determined as in Figure 3A. The zero minute time point was taken immediately after each treatment. Error bars represent SD (n = 2 experiments). (B) Boxplots of intracellular pH after hyposmotic shock as described in Figure 5E. Cells were treated with hyperosmotic shock media for 10 min before ratiometric imaging. Values from two biological replicates were pooled and compiled into boxplots. Whiskers represent the minimum and maximum respectively. Outliers (black circles) are defined as points further from the quartile-box than 1.5 times the length of the quartile-box. (C) Log-log MSD plot of the pLacO plasmid. Cells were treated as described in Figure 5E. (D) Log-log MSD plot of the FBA1 mRNP. Cells were treated as described in Figure 5E.

DOI: http://dx.doi.org/10.7554/eLife.09376.016