Effects of MF286 on in vitro SNARE binding and GLUT4 translocation. (A) Blood glucose concentrations during an IPGTT after injections of PBS or MF286. The data were obtained from four PBS-injected mice and three MF286-injected mice. (B) Plasma insulin concentrations in PBS- or MF286-injected mice without (glucose −) or with (glucose +) glucose stimulation. The data are representative of three independent experiments. (C) Percentages of initial blood glucose concentration during an ITT after injection of PBS or MF286. The data are representative of three independent experiments. (D) In vitro SNARE complex formation among GST–syntaxin1A (Stx1A), His-VAMP2, and His-SNAP23 (left) or GST-Stx1A, His-VAMP2, and His-SNAP25 (right) with or without MF286. (E) In vitro SNARE complex formation among GST-Stx4, His-VAMP2, and His-SNAP23 (left) or GST-Stx4, His-VAMP2, and His-SNAP25 (right) with or without MF286. (F) GLUT4 staining in 3T3-L1 adipocytes before or after insulin stimulation. Bar, 20 µm. (G) Insulin-stimulated glucose uptake in 3T3-L1 adipocytes with or without MF286 treatment. The data are representative of four independent experiments. (H) CCK-stimulated amylase secretion from isolated acinar cells treated with PBS, 37.5 µM MF286, and 300 nM tetanus toxin (TeTx). Tetanus toxin, which cleaves VAMP2 in pancreatic ZGs and inhibits enzyme secretion, was used as a positive control. The data are representative of four independent experiments. Data are mean ± SEM. Significance was calculated by the two-tailed paired Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.