Supplementary data

Files in this Data Supplement:

• Supplementary figure 1 - Gene expression profile of established Men1Τ/Τ ES cell lines. Gene expression of Oct3/4, Nanog and Sox1 as demonstrated by RT-PCR reaction with mRNA isolated from Men1 wild type (Men1+/+) and Men1 null (Men1Τ/Τ) ES cell lines, using mRNA isolated from mouse neurospheres as positive control for Sox1, and Hprt as internal mRNA loading control. RT0: PCR using mRNA preparation without reverse transcriptase.
• Supplementary figure 2 - MENIN is specifically required for normal development of the endocrine pancreas in pancreatic bud cultures. Representative whole serial images of pancreatic bud cultures from Men1 wild type (Men1+/+) and null mutant (Men1Δ/Δ) embryos co-stained with antibodies against insulin (red) and carboxy-peptidase A (CPA, green). Note that the number of sections of each cultured bud often varies. The scale bars are 100µm.
• Supplementary figure 3 - MENIN is required for the development of both α and β-Cells in pancreatic bud cultures. Representative whole serial images of pancreatic bud cultures from Men1wild type (Men1+/+) and null mutant (Men1Δ/Δ) embryos co-stained with antibodies against insulin (red) and glucagon (green). The scale bars are 100µm.
• Supplementary figure 4 - Absence of insulin- and glucagon-secreting cells on large cystic structures in cultured Men1 null pancreatic buds. Representative images of Men1 wild type (A and B) or null(C and D) pancreatic bud cultures at day 7 co-stained either with antibodies against insulin (red) and carboxy-peptidase A (CPA, green), or with insulin (red) and glucagon (green). Arrows show the edge of cyst.
• Supplementary figure 5 - Disruption of the Men1 gene results in loss of Ngn3-expressing cells in day 2 pancreatic bud cultures. Representative serial images of in situ hybridization for Ngn3 expression in a two-day cultured Men1 null(Men1Δ/Δ) and a control (Men1+/+) cultured bud. The scale bars are 100µm.