Figure 2.
During hypoosmotic swelling, tricalbin mutants lose cell integrity. (A–C) Using time-lapse imaging, wild type and mutant strains expressing pHluorin-mCherry (SEY6210, MTY2, and MTY18 containing pMB517) were observed during exposure to hypoosmotic conditions (1 M sorbitol > 0 M sorbitol). The resulting movies were analyzed for cell surface changes and changes in cytoplasmic pH (pHluorin/mCherry ratio). Downward arrows in C mark cell lysis events. (D) Quantification of the data obtained by the time-lapse imaging presented in box-and-whisker plots (line indicates the mean). The first panel in C explains the measured parameters. (E) Wild type (SEY6210) and tricalbin mutant (MTY18) cells were either moved to the same sorbitol-containing medium (1 M sorbitol > 1 M sorbitol) or exposed to a hypoosmotic shock (1 M sorbitol > 0 M sorbitol) at 30°C. 5 min after treatment, the cells were plated and colonies were counted after 2 days of growth.

During hypoosmotic swelling, tricalbin mutants lose cell integrity. (A–C) Using time-lapse imaging, wild type and mutant strains expressing pHluorin-mCherry (SEY6210, MTY2, and MTY18 containing pMB517) were observed during exposure to hypoosmotic conditions (1 M sorbitol > 0 M sorbitol). The resulting movies were analyzed for cell surface changes and changes in cytoplasmic pH (pHluorin/mCherry ratio). Downward arrows in C mark cell lysis events. (D) Quantification of the data obtained by the time-lapse imaging presented in box-and-whisker plots (line indicates the mean). The first panel in C explains the measured parameters. (E) Wild type (SEY6210) and tricalbin mutant (MTY18) cells were either moved to the same sorbitol-containing medium (1 M sorbitol > 1 M sorbitol) or exposed to a hypoosmotic shock (1 M sorbitol > 0 M sorbitol) at 30°C. 5 min after treatment, the cells were plated and colonies were counted after 2 days of growth.

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