Fig. 3.

Dose-dependency of mitochondria and Ca²⁺ concentrations for the CRC/Mitochondrial swelling assay. (a) and (b) Varying amounts of mitochondria (0.5, 1, 2, 4 mg) subjected to the CRC/swelling assay using the same additions of 20 μM Ca²⁺. Representative traces demonstrate that increasing the concentration of mitochondria in the cuvette correlates with an increase in total Ca²⁺ required to induce MPTP opening. (c) Quantification of figure a showing that the CRC remains constant although increasing amounts of mitochondria were used (n = 3). (d) and (e) 2 mg of mitochondria subjected to the CRC/swelling assay using increasing concentrations of Ca²⁺ additions (10, 20, 40 μM). Representative traces show that MPTP opening is achieved sooner with higher additions of Ca²⁺, demonstrating the dose-dependent nature of CRC assay. (f) Quantification of figure d showing that the CRC remains constant although increasing amounts of Ca²⁺ were used (n = 3) error bars (S.E.M)