Exosomal transfer of miR-375-3p from pancreatic β cells to hepatocytes impairs hepatic glycogenesis via Rbpj repression

Figure 6 miR-375-3p regulates hepatocyte glycogenesis via Rbpj. A: Schematic diagrams illustrating the miR-375-3p binding site in the Rbpj 3'-untranslated region (UTR) predicted by TargetScan. Hep1-6 cells were transfected with a miR-375-3p inhibitor or control inhibitor for 48 hours; B-E: Western blotting of AKT/GSK signaling and protein level of Rbpj; F: Detection of glycogen content. Hep1-6 cells were cotransfected with miR-375-3p mimics and Rbpj expression plasmid or empty vector for 48 hours; G-J: Western blotting of AKT/GSK signaling and the protein level of Rbpj; K: Detection of the glycogen content; L-N: Quantification of Rbpj protein (L and M) and mRNA (N) levels in Hep1-6 cells transfected with miR-375-3p mimics or control mimics; O-Q: Protein and RNA levels of Rbpj in the livers of high-fat diet-fed mice and chow diet-fed mice; R-U: Protein and RNA levels of Rbpj in Hep1-6 cells treated with exosomes secreted from MIN-6 cells after low- or high-glucose stimulation (R and S) or with miR-375-3p inhibitor (T and U); V: Schematic diagrams of the mutated miR-375-3p binding site in the Rbpj 3'-UTR; W and X: Hep1-6 cells were cotransfected with miR-375-3p mimics and pmirGLO-Rbpj-WT/MUT plasmids, followed by luciferase reporter assay. The experiments were repeated three times. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. Western blot raw data in Supplementary material.