ose induced synthesis of insulin The glucose induced synthesis of insulin in the mice hepatocytes due to the expression of both proinsulin genes I and II in mice was determined by incubating the GLS in HEPES buffer with different concentrations of glucose for 30 min at 37C. After incubation the extracted nucleic acids which contained 
insulin-mRNA was translated in vitro by using plant ribosomal particles as described before and synthesized insulin was subsequently quantitated by ELISA by using both anti-insulin antibody and commercial insulin antibody. The synthesis of insulin in the assay mixture was confirmed by bioassay of the hormone as described before. The expression of both proinsulin genes I and II were determined by cDNA preparation as described before. 4 Glucose purchase Halofuginone Transporter-4 in Mice Hepatocytes doi: 10.1371/journal.pone.0081935.g002 Statistical Methods The results shown are mean standard deviation of at least 5 experiments using 5 different animals, each carried out in triplicate. The significance of the results was analyzed by Student’s t test. Significance was considered to be significant. Results Effect of glucose on the activation of nitric oxide synthase in liver cell membrane from adult mice As the treatment of islets of Langerhans with glucose has been reported to result in the stimulation of NO synthesis, to determine the possibility of the occurrence of the glucose induced synthesis of NO in the liver cells, the liver membrane preparation was treated with different amounts of glucose to determine the possible occurrence of the glucose activated nitric oxide synthase in the membrane preparation. The treatment of the liver cell membrane preparation with different amounts of glucose as indicated was found to result in the increased synthesis of NO, and at 0.02M glucose the liver membrane GANOS activity was found to be maximally stimulated. Addition of 0.1mM NAME, an inhibitor of NO synthesis, was found to completely nullify the stimulatory effect of the added glucose on GANOS at all concentrations of glucose used in the synthesis of NO in the assay mixture. 5 Glucose Transporter-4 in Mice Hepatocytes doi: 10.1371/journal.pone.0081935.g003 Lineweaver-Burk plot of the stimulation of GANOS by glucose in the liver membrane preparation Lineweaver-Burk plot of the activity of GANOS in the liver membrane preparation, in the presence of 0.02M glucose, indicated that the basal GANOS activity of the membrane preparation that indicated NO production by GANOS was stimulated due to the addition of 0.02M glucose in the reaction mixture . Effect of glucose induced NO synthesis in the GLS in the uptake of the sugar by the suspension The treatment of GLS with different amounts of glucose not only resulted in the increased synthesis of NO by the suspension as described, but was also found that the increased synthesis of NO resulted in the increased uptake of glucose by the 14579267 suspension, and at 120 min, the uptake of glucose was maximally achieved. The addition of 0.1mM NAME to the reaction mixture that completely inhibited NO synthesis was found to simultaneously abolish the glucose induced glucose transport in 15102954 the GLS 6 Glucose Transporter-4 in Mice Hepatocytes doi: 10.1371/journal.pone.0081935.g004 The glucose induced translocation of Glut-4 in the liver cells membrane The results described in immunohistochemical technique could not be demonstrated. The synthesis of Glut-4 in GLS in the presence of glucose As glucose was found to increas
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