Fig. 5. Determination of the ANT content in isolated mitochondria by CAT titration. (A) CAT titre of respiration. State 3 respiration was titrated by successive additions of CAT to achieve state 4 respiration. ANT content was measured as the CAT titre where the steepest slope in the titration crosses the state 4 rate (broken lines). Results of a single representative determination are shown for liver mitochondria from fed cane toads acclimated to 30°C. (B,C) ANT contents of liver and skeletal muscle mitochondria of all experimental conditions measured by CAT titre (B, WA cane toads; C, CA cane toads). ANT content is about five times lower in liver mitochondria compared to skeletal muscle (P<0.05, two-way ANOVA.) but neither the temperature nor the nutritional status have an effect. Values are means ± s.e.m. from 4 independent preparations.^*P<0.05 (two-way ANOVA).