Assessing Cell Death by Propidium Iodide

Spotting onto drug-free medium was performed to evaluate cidality as previously described (69 (link)). Plates were incubated at 30°C and photographed after 24 h. Propidium iodide staining was used to visualize cell death. Cells were subcultured from a saturated overnight culture to an OD₆₀₀ of 0.1 in YPD medium and grown for 18 h in the absence or presence of 50 μM CMLD010515. Cells were pelleted and resuspended in phosphate-buffered saline (PBS) with 1 μg/ml propidium iodide. Cells were imaged by differential interference contrast (DIC) microscopy and by the use of the DsRed channel on a Zeiss Axio Imager.MI microscope (Carl Zeiss) at the same exposure time.