Effects of biotin deficiency on pancreatic islet morphology, insulin sensitivity and glucose homeostasis Elena Larrieta, Maria Luisa Lazo de la Vega-Monroy,

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Effects of biotin deficiency on pancreatic islet morphology, insulin sensitivity and glucose homeostasis Elena Larrieta, Maria Luisa Lazo de la Vega-Monroy, Paz Vital, Asdrubal Aguilera, Michael S. German, Mohammed El Hafidi, Cristina Fernandez-Mejia Journal of Nutritional Biochemistry Volume 23, Issue 4, Pages 392-399 (April 2012) DOI: 10.1016/j.jnutbio.2011.01.003 Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 1 Effect of biotin deficiency on biotinylated pyruvate carboxylase. (A) Western blot analysis of biotinylated pyruvate carboxylase abundance in the liver of mice fed a control and biotin-deficient diet for 8 weeks. Control mice: lines 1–3; biotin-deficient mice: lines 4–6. (B) Quantification of biotinylated pyruvate carboxylase abundance in the liver. White bars: control group; dashed bars: biotin-deficient group. Data are mean±S.E. of values from three mice of each group. ⁎P<.05 vs. control mice. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 2 Effect of biotin deficiency on glucose tolerance. Three-week-old male BALB/cAnN Hsd mice were fed either a biotin-deficient diet or a control-diet during 8 weeks. (A) Intraperitoneal glucose tolerance test. (B) Relative glucose values in intraperitoneal glucose tolerance test. Open circles: control group; closed circles: biotin-deficient group. Data show the mean±S.E. of values from 5–10 mice of each group. ⁎Indicates significant differences (P<.05) between groups. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 3 Effect of biotin deficiency on insulin sensitivity. Relative glucose values in intraperitoneal insulin tolerance test. Open circles: control group; closed circles: biotin-deficient group. Data are the mean±S.E. of values from 5–10 mice of each group. ⁎Indicates significant differences (P<.05) between groups. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 4 Effect of biotin deficiency on hepatic insulin receptor protein mass. (A) Representative Western blot analysis of insulin receptor protein mass. (B) Quantification of insulin receptor protein mass abundance. White bars: control group; dashed bars: deficient-diet group. Graph data are means±S.E. of values from 5–10 mice of each group. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 5 Effect of biotin deficiency on phosphorylated and nonphosphorylated Akt/PKB. (A) Total Akt. (B) Phosphorylated Akt. White bars: control group; dashed bars: deficient-diet group. Graph data show the mean±S.E. of values from four mice of each group. ⁎Indicates significant differences (P<.05) between groups. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 6 Effect of biotin deficiency on pancreatic morphology. (A) Islet architecture immunostaining analysis: amplification, 40×. Calibration bar, 50 μm. Insulin (green); glucagon (blue). (B) Percent of islets with α-cells present in the center of the islet. Data show the mean±S.E. of values from 10 mice of each group. ⁎Indicates significant differences (P<.05) between groups. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 7 Pancreatic morphometry analysis. (A) Percentage of islet area per pancreas area. (B) Number of islets per pancreas area. (C) Frequency distribution of islet size. (D) Glucagon-positive area and insulin-positive area per islet area. Data show the mean±S.E. of values from three mice of each group. ⁎P<.05 vs. the corresponding value for control mice. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions

Fig. 8 Glucagon mRNA abundance and secretion. (A) Relative mRNA abundance of glucagon. Glucagon and 18s rRNA were quantified by real-time PCR. Data are expressed as fold change relative to control group. Values show the mean±S.E.M. of four to five pooled samples of 200 islets. (B) Glucagon secretion. Graph data are the mean±S.E. of values from five independent experiments. ⁎Indicates significant differences (P<.05) between groups. Journal of Nutritional Biochemistry 2012 23, 392-399DOI: (10.1016/j.jnutbio.2011.01.003) Copyright © 2012 Elsevier Inc. Terms and Conditions