Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Supplemental Table 1 - The oligo sequences of primers used both for genotyping and quantitative PCR. The first three primers were used for genotyping, the following primers were used for SYBR green-based QPCR and the last five fluorogenic probes were used for TaqMan assays.
• Supplemental Table 2 - Individual measurement for insulin content performed on isolated islets from littermate pairs of GPR39+/+ and GPR39 -/- mice, which have been handled in parallel experimentally. The period of culture differed form 3 to 5 days as indicated in the last column.
• Supplemental Figure 1 - Verification of the specificity of the anti-sera against GPR39-1a. HEK293 cells transiently transfected empty vector/mock transfection (left panel) and GPR39-1a (right panel).
• Supplemental Figure 2 - QPCR analysis of the expression of Gpr39 in rat liver, white adipose tissue (WAT), pancreas, and isolated islets as well as in a rat insulinoma cell line, 12C3IN, and a corresponding glucagonoma cell line, 12C3AN (Madsen,O.D., Karlsen,C., Nielsen,E., Lund,K., Kofod,H., Welinder,B., Rehfeld,J.F., Larsson,L.I., Steiner,D.F., Holst,J.J. et al 1993. The dissociation of tumor-induced weight loss from hypoglycemia in a transplantable pluripotent rat islet tumor results in the segregation of stable alpha- and beta-cell tumor phenotypes. Endocrinology 133:2022-2030).
• Supplemental Figure 3 - Targeted disruption of the GPR39 gene. (A) Structure of the neomycin resistance containing targeting construct, the wild type GPR39-1a locus and the recombinant locus. Grey boxes represent untranslated regions. The green boxes represent the open reading frame of GPR39 whereas the red box represents the open reading frame for the neomycin resistance gene. (B) A representative gel showing the genotyping. (C) Relative expression of GPR39 in the pancreas from GPR39 (+/+), (+/-) and (-/-) mice.
• Supplemental Figure 4 - Oral glucose tolerance tests (OGTT) performed in GPR39+/- (black triangle and black line) and GPR39 +/+ (empty circle and dottedīline), respectively, at 11-12 weeks of age; glucose was measured in tail blood.
• Supplemental Figure 5 - Perifusion of islets isolated from Gpr39(-/-) mice (black circles and black line) and wild-type mice (empty circles and dotted line) exposed to step-wise increasing concentrations of glucose. The age of the mice in both groups were 20 weeks. Data are mean ± s.e.m.. n=4
• Supplemental Figure 6 - Perifusion of islets isolated from Gpr39(-/-) mice (black circles and black line) and wild-type mice (emptycircles and dotted line) exposed to step-wise increasing concentrations of glucose. The age of the mice in both groups were 16 weeks. Data are a representative experiment of n=4