The glucose-6-phosphate dehydrogenase from mouse liver is fully inhibited in vitro by physiological concentrations of NADPH. This suggests that the oxidative phase of the pentose phosphate pathway requires some deinhibitory system. In order to investigate regulation of the pentose phosphate pathway, various parameters (intermediate concentrations, mass-action ratios of reactions, etc.) were measured in liver from control mice and from meal-fed mice. Assays were also carried out to detect any molecules causing the reverse of glucose-6-phosphate dehydrogenase inhibition by NADPH. The liver of meal-fed mice show greater glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities. They also had greater concentrations of several metabolic intermediates and triglycerides than the control animals (P < 0.001). These results prove that the diet increases the flow of the pentose phosphate pathway in a lipogenic sense. The glutathione reductase does not change with the diet, suggesting that this enzyme does not participate in the modulating process. Unlike rat liver, no molecules causing the reverse of glucose-6-phosphate dehydrogenase inhibition by NADPH were detected. These data suggest that the increase of flow of the pentose phosphate pathway during lipogenesis is obtained by an increase in enzyme synthesis.