Intravenous infections of minute doses of triiodothyronine were administered to thyroidectomized rats 30 minutes before they were killed. Hepatic mitochondria were isolated rapidly and formation of adenosine triphosphate and consumption of oxygen were assessed by a 2-minute incubation. Hormone injection enhanced formation of adenosine triphosphate 114 to 217 percent over control values, with a proportionate increase in consumption of oxygen. The ratio of phosphate to oxygen was about 2.0, signifying tightly coupled oxidative phosphorylation. Stimulation was not abolished by injection of cycloheximide, puromycin, actinomycin D, or chloramphenicol 1 hour before the rats were killed. This signifies direct mitochondrial stimulation by triiodothyronine in the absence of protein synthesis.