Rotenone was applied to the cells as following concentrations: 0nM, 50nM, 100nM, 200nM, 400nM and 800nM. The cells have been monitored by IncuCyte FLR imaging system (cell confluence as the marker). Also, I applied two assays, CellTox Green Cytotoxicity Assay and Essen CellPlayerTM 96- Well Kinetic Caspase-3/7 Apoptosis Assay, to the cells in order to detect apoptotic cells/signs of cytotoxicity (green fluorescence) but neither one of them gave me promising results. Any ideas why these cells aren't dying because of rotenone?
Hi Marie, I had the same problem with rotenone this past while, but on PC12 cells. How long did you expose your cells to rotenone? I decided on increasing the rotenone concentration to a maximum of 10 uM, treating them for both 24- and 48 hours. I chose the sulforhodamine B assay as rotenone targets mitochondrial complex I and could affect mitochondrial-dependent cytotoxicity assay results (MTT, CeltitreBlue and CeltitreGlo etc.). I treated the PC12s today and repeating the experiment on SH-SY5Y cells next week. All the best!
We exposed them to rotenone 24-72 hours, and monitored them with IncuCyte during the whole timeperiod. Is the sulforhodamine B assay only for end-point analysis?
I wanna treat PC12 with Rot, and I wanna to know if you have the promising results. If yes, can you tell me the dosage and time you used?
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