To begin, seed and culture wild type medulloblastoma cell lines in a six well plate. After 24 hours, add the appropriate ferroptosis inducing agent to the cells and incubate the plate at 37 degrees Celsius with 5%carbon dioxide. After six hours of treatment, observe the cells under a light microscope for cell rounding.
Using an aspirator, remove the media and floating cells from the plate under a Laminar airflow. To wash the cells, add two milliliters of PBS, swirl gently and aspirate the PBS out. Next, add two milliliters of the probe working solution and incubate the plate in the dark.
Light, microscopic images of the DAOY wild type cells treated with one micromolar of Erastin showed round bubbling cells indicating a ferroptotic phenotype.
