1. Volume 18, Issue 13, Pages 3192-3203 (March 2017)
Pancreatic α Cell-Derived Glucagon-Related Peptides Are Required for β Cell Adaptation and Glucose Homeostasis 
Shuyang Traub, Daniel T. Meier, Friederike Schulze, Erez Dror, Thierry M. Nordmann, Nicole Goetz, Norina Koch, Elise Dalmas, Marc Stawiski, Valmir Makshana, Fabrizio Thorel, Pedro L. Herrera, Marianne Böni-Schnetzler, Marc Y. Donath 
Cell Reports 
Volume 18, Issue 13, Pages (March 2017)
DOI: /j.celrep
Copyright © 2017 The Author(s) Terms and Conditions

2. Cell Reports 2017 18, 3192-3203DOI: (10.1016/j.celrep.2017.03.005)
Copyright © 2017 The Author(s) Terms and Conditions

3. Figure 1
DT Injection in GluDTR Mice Leads to Massive and Long-Lasting α Cell Ablation
(A) Immunohistochemical staining of pancreatic sections from GluDTR and wild-type (WT) mice 63 weeks after diphtheria toxin (DT) injection stained for insulin (green) and glucagon (red). Scale bar, 50 μm.
(B) Gcg and Ins2 mRNA expression in isolated islets from GluDTR and WT mice 6 weeks after DT injection. n = 8.
(C) Glucagon und active GLP-1 content of isolated islets from 13-week-old GluDTR and WT mice. n = 4.
(D) Immunohistochemical staining of transversal colon sections from 31-week-old GluDTR and WT mice stained for GLP-1 (red). White arrowheads indicate GLP-1-positive L cells. Scale bar, 25 μm.
(E) Active GLP-1 content in terminal ileum of 54-week-old GluDTR and WT mice. n = 6 and 7.
(F) Body weight development of GluDTR versus WT mice after DT injection. n = 6.
(G) Fasting plasma glucagon levels in 30-week-old GluDTR and WT mice. n = 6.
(H) Fasting plasma active GLP-1 levels in GluDTR and WT mice. n = 16 and 12.
n indicates the number of independent experiments or number of animals respectively, and error bars represent SEM. ∗p < See also Figure S1.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

4. Figure 2
α Cell-Derived GLP-1 Is Required for Normal i.p. Glucose Tolerance during Aging
(A and B) Intraperitoneal (i.p.) glucose tolerance test (A) and corresponding insulin levels (B) in GluDTR and WT mice at the age of 12 weeks. n = 13.
(C and D) i.p. glucose tolerance test (C) and corresponding insulin levels (D) in GluDTR and WT mice at the age of 28 weeks. n = 20.
(E and F) i.p. glucose tolerance test (E) and corresponding insulin levels (F) in GluDTR and WT mice at the age of 30–32 weeks. Sitagliptin (25 mg/kg body weight) was i.p. injected 30 min prior to glucose injection. n = 13 and 12.
(G and H) i.p. glucose tolerance test (G) and corresponding insulin levels (H) in GluDTR and WT mice at the age of 32–34 weeks. n = 13 and 12. Glucagon (20 μg/kg body weight) was i.p. injected 1 min prior to glucose injection.
n indicates the number of animals, and error bars represent SEM. ∗p < 0.05, ∗∗p < 0.01, area under the curve (AUC). See also Figure S2.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

5. Figure 3
Oral Glucose Tolerance Is Improved in Young α Cell-Ablated Mice due to Increased Systemic Active GLP-1 Secretion
(A) Oral glucose tolerance test in GluDTR and WT mice at the age of 12 weeks. n = 15 and 13.
(B) Active GLP-1 content in terminal ileum of 8- to 12-week-old GluDTR and WT mice. 1 = ± 24.7 ng/mL. n = 14 and 12.
(C and D) Plasma active GLP-1 concentration (C) and plasma insulin concentration (D) during oral glucose tolerance test in 13-week-old GluDTR and WT mice that were injected with sitagliptin (25 mg/kg body weight) 30 min prior to glucose administration. n = 18 and 12.
(E–G) Blood glucose (E), insulin (F), and active GLP-1 (G) levels during an oral glucose tolerance test with or without glucagon in GluDTR and WT mice at the age of 13–15 weeks that were injected with sitagliptin (25 mg/kg body weight) 30 min prior to glucose administration (n = 11). Glucagon (50 μg/kg body weight) or vehicle control was i.p. injected 1 min prior to glucose administration.
(H and I) Blood glucose (H) and insulin (I) during an oral glucose tolerance test in GluDTR and WT mice at the age of 28 weeks. n = 18 and 13.
(J) Oral glucose tolerance test with or without exendin(9–39) in GluDTR and WT mice at the age of 28 weeks. n = 9, 10, 6, 7. Exendin(9–39) (70 nmol/kg body weight) or vehicle control was intraperitoneally injected 30 min prior to glucose administration.
n indicates the number of animals, and error bars represent SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p <
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

6. Figure 4
α Cell Ablation Protects against HFD-Induced Glucose Intolerance by Increasing Insulin Sensitivity
(A and B) Blood glucose (A) and insulin (B) levels during an i.p. glucose tolerance test in in 27-week-old GluDTR and WT mice following 21 weeks of high-fat diet (HFD) feeding. n = 13 and 11.
(C and D) Blood glucose (C) and insulin (D) levels during an oral glucose tolerance test in GluDTR and WT mice following 22 weeks of HFD feeding. n = 13 and 11.
(E) Insulin tolerance test in GluDTR and WT mice after 23 weeks of HFD feeding. n = 13 and 11.
(F–H) Body weight (F), fasting plasma active GLP-1 (G), and fasting plasma glucagon (H) concentrations in GluDTR and WT mice following 22 weeks of HFD feeding. DT injections were at 5–6 weeks of age and HFD was started at 6–7 weeks of age.
n indicates the number of animals, and error bars represent SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p <
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

7. Figure 5
α Cell-Derived Glucagon-Related Peptides Are Required for Normal GSIS and β Cell Function via cAMP-Mediated β Cell Potentiation
(A) Glucose-stimulated insulin secretion (GSIS) of isolated islets from GluDTR mice that had been injected with DT or saline. n = 3.
(B) GSIS of isolated C57BL/6N mouse islets with or without co-incubation of 100 nM exendin(9–39) during stimulation with 16.7 mM glucose. n = 3.
(C) Relative luminescence of the GLP1R activity luciferase reporter assay. 48 hr after transfection with the GLP1R-Tango vector, HTLA cells were treated with human islet conditioned medium (HI) with or without 100 nM exendin(9–39) or human islet medium containing 100 nM GLP-1 with or without 100 nM exendin(9–39). n = 3.
(D) GSIS and corresponding stimulatory index (fold increase stimulated over basal insulin release) of GluDTR and WT islets with or without co-incubation of 10 nM GLP-1 or 10 nM glucagon during 16.7 mM glucose stimulation. 1 = 7.1 ± 1.5 ng/mL. n = 3.
(E) Cumulative insulin secretion over 24 hr. 1 = ± 87.8 ng/mL. n = 3.
(F) Cellular insulin content of GluDTR and WT islets. 1 = ± 109.5 ng/mL. n = 3.
(G) GSIS and corresponding stimulatory index of GluDTR and WT islets with or without co-incubation of 10 μM forskolin during 16.7 mM glucose stimulation. 1 = 100.0 ± 12.2 ng/mL. n = 3.
n indicates the number of independent experiments, and error bars represent SEM. ∗∗p < 0.01, ∗∗∗p < See also Figure S3.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

8. Figure 6
α Cell-Specific PC1/3 Knockout Results in Decreased Islet Active GLP-1 Secretion and Cell Content
(A) Active GLP-1, glucagon and insulin release over 24 hr from isolated αPcsk1−/− and control islets. n = 3.
(B) Active GLP-1, glucagon and insulin content of isolated αPcsk1−/− and control islets. n = 3.
(C) GSIS and corresponding stimulatory index (fold increase of stimulated over basal insulin release) of isolated αPcsk1−/− and control islets with or without co-incubation of 10 nM GIP during 11.1 mM glucose stimulation. 1 = 12.5 ± 2.4 ng/mL. n = 3.
(D) Body weight development of αPcsk1−/− and control mice. n = 9 and 12.
(E) Fasting plasma active GLP-1 concentration in 15-week-old αPcsk1−/− and control mice. n = 16.
(F) RNA profile of isolated islet from αPcsk1−/− and control mice.
n indicates the number of independent experiments or the number of animals, and error bars represent SEM. ∗p < 0.05, ∗∗p < See also Figure S4.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions

9. Figure 7
i.p. Glucose Tolerance Is Improved in Chow-Diet-Fed α Cell-Specific PC1/3 Knockout Mice and Is Impaired during Metabolic Stress
(A and B) Blood glucose (A) and corresponding insulin (B) levels during an i.p. glucose tolerance test in αPcsk1−/− and control mice at the age of 11 weeks. n = 16.
(C and D) Blood glucose (C) and insulin (D) levels during an oral glucose tolerance test in αPcsk1−/− and control mice at the age of 12 weeks. n = 9.
(E–G) Body weight (E), fasting plasma active GLP-1 concentration (F), and active GLP-1 content (G) in terminal ileum of 15-week-old HFD/streptozotocin (STZ)-treated αPcsk1−/− and control mice. n = 20 and 24.
(H and I) Blood glucose (H) and corresponding insulin (I) levels during an i.p. glucose tolerance test in HFD/STZ-treated αPcsk1−/− and control mice at the age of 14 weeks. n = 26 and 31.
(J and K) Blood glucose (J) and corresponding insulin (K) levels during an i.p. glucose tolerance test in 15-week-old αPcsk1−/− and control mice that were injected with sitagliptin (25 mg/kg body weight) 30 min prior to glucose administration to stabilize active GLP-1. n = 15 and 17.
n indicates the number of animals, and error bars represent SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < See also Figure S5.
Cell Reports  , DOI: ( /j.celrep )
Copyright © 2017 The Author(s) Terms and Conditions