Case Studies

Mitochondrial toxicity assessment

Mitochondrial toxicity assays provide a means to measure mitochondrial dysfunction due to the toxic effect of a test compound. Our robust and sensitive assay setup can identify or rule out potential toxicity caused by mitochondrial impairment.

ATP Synthesis Assay

Figure-A: Measuring ATP generation in the presence of glucose (glucose media) and absence of glucose (galactose media). Cells treated with tool compounds and ATP levels measured using ATPlite luminescence readout. Percent (%) ATP levels calculated by normalisation to vehicle response. Culturing cells in glucose media favour glycolysis over oxidative phosphorylation for producing ATP. Cells cultured in absence of glucose, forces cells to generate ATP by oxidative phosphorylation there by we can identify compounds which cause mitochondrial impairment.

Membrane Potential Assay

Figure-B: Mitochondrial Membrane Potential (MMP) assay utilises JC-10 dye for measuring mitochondrial depolarization in cells. JC-10 is a cationic dye accumulates in energised mitochondria. Cells treated with tool compounds. At high membrane potentials, the dye aggregates and at low membrane potentials the dye exists as a monomer. Ratio of monomer/aggregate JC-10 fluorescence intensity was measured as an indicator of mitochondrial impairment.

Identifying potential mitochondrial toxins as early as possible in drug discovery is crucial. Here we demonstrate sensitive assay design approach as an indicator to assess drug-induced mitochondrial toxicity. Compound IC50  was derived from Dose Response curves in ATP synthesis assay and Mitochondrial Membrane Potential (MMP) assay.

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