IF1 (ATPIF1) is a nuclear DNA-encoded mitochondrial protein whose activity is inhibition of the F1Fo-ATP synthase to control ATP production.IF1 activity remains unknown in the regulation of GLP-1 activity.In this study,IF1 was examined in the diet-induced obese mice using the gene knockout(IfI-KO) mice.The mice gained more body weight on a high fat diet without a change in food intake.Insulin tolerance was impaired,but the oral glucose tolerance was improved through an increase in GLP-1 secretion.The KO mice exhibited an improved intestine structure,mitochondrial superstructure,enhanced mitophagy,reduced apoptosis and decreased adenine nucleotide translocase 2 (ANT2) protein in the intestinal epithelial cells together with preserved gut microbiota.The data suggest that GLP-1 secretion was enhanced in the obese If1-KO mice to preserve glucose tolerance through a signaling pathway of ANT2/mitochondria/L-cells/GLP-1/insulin.IF1 is a potential mitochondrial target for induc-tion of GLP-1 secretion in L-cells.