Differentiation of cells expanded from human islet cells into adipocytes and osteocytes.

<p>A, Islet cells at the indicated passage number were incubated in Lonza induction medium and stained with Oil Red O for adipocytes and Alizarin Red for osteocytes. Human BM-MSC served as positive control. Bar = 100 µm. B, Islet cells at passage 5 and BM-MSC were incubated in Lonza adipogenesis induction medium and stained with antibodies to eGFP and FABP4. Nuclei were stained blue with DAPI. Bar = 30 µm. The eGFP⁺ cells shown do not stain for FABP4. The single FABP4⁺ cell shown is not eGFP⁺. C, Quantitation of the staining in B, based on counting >500 cells in cultures derived from each donor. Data represent percent of FABP4⁺ among eGFP⁺ cells (green bars) and among all the cells (black bars) and are mean±SD (n = 6 donors for islet cells and 2 donors for BM-MSC). p = 4.18E-14.</p

Inducible labeling of human islet cells.

<p>A, Schematic representation of the two lentivirus vectors. B, Labeling efficiency and leakiness. Adult human islet cells from 5 donors were infected one day after plating with the reporter virus alone, or with both viruses, and cultured overnight in the absence or presence of tamoxifen. Five days later 10⁴ cells from each donor were analyzed by flow cytometry for eGFP expression. Data are mean±SD (n = 5).</p

Expression of mesenchymal genes in eGFP⁺ cells.

<p>Cells at passage 4 were stained with antibodies to eGFP and the indicated mesenchymal marker. Nuclei were stained blue with DAPI. Bar = 20 µm. The percentages indicate the fraction of cells positive for each mesenchymal marker among eGFP⁺ cells (green digits) and among all the cells (white digits). Data are mean±SD of >400 cells scored from each donor (n = 3 donors).</p

Changes in expression of epithelial and mesenchymal genes in human islet cells during the first 3 weeks in culture.

<p>RNA was extracted from cells at the indicated passage number (each passage is equivalent to one week) and analyzed by qRT-PCR. A, analysis of epithelial genes. B, analysis of mesenchymal genes. Data represent relative quantification (RQ) (compared to passage 0) and are mean±SE (n = 6 donors). *p<0.05; ** p<0.005; ***p<0.0005 (compared to passage 0).</p

Changes in expression of epithelial and mesenchymal genes in sorted eGFP⁺ and eGFP⁻ cells following 2 weeks in culture.

<p>RNA was extracted from cells at the indicated passage number and analyzed by qRT-PCR. A, analysis of epithelial genes. B, analysis of mesenchymal genes. Data represent relative quantification (RQ) (compared to passage 0) and are mean±SE (n = 3 donors). *p<0.05; ** p<0.005.</p

Association results for the <i>IL2RA</i> region and CpG −373.

<p> Manhattan plot represented p-values of Wald test based on linear regression model (for each SNP, association test performed in plink which compares the quantitative phenotype means for three genotypes). The red horizontal line represents the significance threshold of P = 4.7×10⁻⁴.</p

Main characteristics and CpG methylation levels in the <i>IL2RA</i> promoter of T1D patients and age-matched non-diabetic controls.

<p>Results are expressed as mean ± sd.</p

Schematic representation of DNA methylation levels in the proximal promoter of the <i>IL2RA</i> gene.

<p>Tissues come from different non-diabetic controls: WBC (n = 286), liver (n = 7), peritoneum (n = 8), thymus (n = 16) and Langerhans islets (n = 7), regulatory T cells (n = 8).</p