Figure 8.

Deletion of wat1 gene results in increased ROS production and cell death. Midlog phase culture of wild-type and wat1-deleted cells were incubated for 90 min at 25° in the presence of DCDHFDA to detect ROS. For the 36° experiment, the midlog phase cultures were shifted at 36° for 3 hr, then incubated with media containing DCDHFDA for 90 min at 36°. The cells were harvested, washed, and resuspended in buffer containing PI to detect dead cells. This experiment was performed three times with similar results. DCDHFDA, 2′,7′-dichlorodihydrofluorescein diacetate; H₂O₂, hydrogen peroxide; PI, propidium iodide; ROS, reactive oxygen species.