Figure 3 | Scientific Reports

Figure 3

From: Identification of ER-000444793, a Cyclophilin D-independent inhibitor of mitochondrial permeability transition, using a high-throughput screen in cryopreserved mitochondria

Figure 3

Identification of ER-000444793 from a high throughput screen using freeze-thawed mitochondria.

Rat liver mitochondria (2 mg protein ml−1), energised using succinate (10 mM) and rotenone (1 μM) were exposed to a CaCl2 bolus (150 μM). Absorbance (540 nm) was measured after 20 minutes. (a) Graphical representation of the metrics from the 50,000 compound screen identifying the ‘hit’ compound cut-off >20% inhibition. (b,c) ER-000444793; N-(2-benzylphenyl)-2-oxo-1H-quinoline-4-carboxamide, was identified from the screen and dose-dependently inhibits Ca2+ induced mitochondrial swelling. (d) Using the same mitochondrial preparations, CsA also delayed pore opening in response to the Ca2+ bolus. Results are expressed as % inhibition, normalised to DMSO (0% inhibition) and CsA (5 μM; 100% inhibition). Data are expressed as means (±s.d.) of at least three independent experiments. Curve fitting used a 4-parameter logistic equation (GraphPad Prism). Abbreviations: IA; inactive, CsA; cyclosporin A.

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