My treatment decreases the level of ROS, but the cells are still continue dying (MTT assay).
I know an example where the ROS decrease, and myeloid cells die because of the presence of IL-2.
good luck!
Are there examples in cardiomyocytes in culture? And in others types of cells in culture. Thank
Concerning the original question, I think that is possible and reasonable. I would need the details about the MTT assay that you use, but it is reasonable that ROS decrease and the cells dye yet.
Any cell death inducer which interferes with fundamental cell functions (i.e. nutrient deprivation, transcription block, translation block, ATP synthesis block, forced open/close of ion/water channels) will eventually kill the cell.
Mainly, the MTT assay measures the mitochondrial red/ox state of your cells. Therefore, if your compound affects the ROS generation it could be posible that your compound might also affect the mitochondrial red/ox state of your cells. To test this you could count death and lives cells using other methods like tripan blue staining.
Other possibility is that your ROS inhibitor is toxic.
I hope you figure out what is happening
Good luck
In many of our studies screening for ROS scavengers, we find no direct or strong correlation between cell death, cell viability, MMP etc. with ROS activity. Often we find an inverse correlation i.e. decreasing ROS activity to a certain point may decrease cell viability significantly. This possibly suggests the complex role of ROS in signal transduction/cellular signalling pathways and not merely a by product of oxidative metabolism . Best wishes!
ROS levels don't necessarily correlate with apoptosis or pre-apoptotic changes in cells. Though often a clear marker of stress and organelle dysfunction, ROS levels can also mark activation of stress signaling during adaptation to the stressor.
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