Fig. 5
Medium osmotic stress affects cell cycle progression and transcription. a Fraction of cells in each cell cycle phases before and during exposure to osmotic stress. Numerical values in Supplementary Fig. S6. The cells were assigned to each phase using the markers previously described in Fig. 1 (n > 1500 cells per time point). b Fluorescence microscopy images depicting a fraction of cells that experience a loss of synchrony between DNA replication and budding under osmotic stress. Red arrows point to cells containing two separated spindle pole bodies without bud. The image is an overlay of bright field image, DAPI-stained nucleus (in blue) and mTurquoise tagged Spc42 part of the spindle pole body (in green). Scale bar represents 5 μm. (c) and (d) n > 400 cells per genes and time points. Raw data in Supplementary Table S1 and Biological Replicates in Supplementary Information. c Changes in mRNA levels shown as mean ± SEM (dots and full lines) and transcription activity represented by percentage of cells with a transcription site (TS) (dots and dotted lines). Plot insets represent a zoom in. d Influence of osmostress on timing of mRNA abundances for SIC1, CLN2, and CLB5 during the cell cycle. Data shown are mean ± SEM. Darkest curves correspond to the shortest incubations in hyperosmotic medium. (e) Simulated changes of timing of mRNA abundances for SIC1, CLB5, and CLN2 during the cell cycle progression upon osmotic stress (n = 2000 cells). Data shown are mean ± SEM. Details of (D) and (E) depicted in Supplementary Fig. S12. In a, c, d, and e 0 min is equivalent to no stress conditions. Lines between data points in c, d, and e are for visualization only
