Male EC-TFEB KO mice were fed a high fat diet (HFD) for 26 weeks. After fasting overnight, the mice were administered insulin (5 U/kg, i.p.). Ten minutes later, tissues were harvested for the analysis of Akt signaling. A-B, the phosphorylation of Akt in the adipose tissue (A) and skeletal muscle (B) of EC-TFEB KO mice was determined by Western blot and quantitatively analyzed (n=3-4/each group). Data in A-B are presented as mean ± SEM; **P <0.01 using 2-way ANOVA followed by Bonferroni test. C, Schematic diagram: endothelial TFEB upregulates insulin receptor substrate 2 (IRS2) at the transcriptional level and increases insulin receptor substrate 1 (IRS1) protein via downregulation of miRNAs, leading to activation of Akt signaling and glucose uptake in ECs. Endothelial TFEB improve systemic glucose tolerance in vivo.