Dear all, I'm having problems determining the proper erastin dose in the SH-SY5Y cell line. Erastin activates ferroptosis and results in ferroptotic cell death. I used a 96-well plate with 4000 cells per well and experimented with various concentrations. I discovered %46 viability when I used 5 uM Erastin twice. Now I'm looking for the dose that results in 70% viability. When I used 2.5 uM Erastin, 80% of the SH-SY5Y cells died. No matter the Erastin dose whether I used 2 uM, 2,5 uM, 3,5 uM, or 5 uM, %80 of the cells died. The control group was fine. Therefore, I don't think there is a vitality issue. Same Erastin, same medium, same plate. Nothing has changed. I don't understand why this is happening. Do you have any suggestions for me to try? Does anyone experience the same problem? Thank you all.
For how long are you treating the cells with erastin? I would suggest to set up the experiment in replicates and use even a lower erastin concentration range and treat the cells for 24, 48, and 72 hrs.
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