Supplementary Figure 1.

Transcription

1 Supplementary Figure 1. Increased expression of cell cycle pathway genes in insulin + Glut2 low cells of STZ-induced diabetic islets. A) random blood glucose measuers of STZ and vehicle treated MIP-GFP transgenic mice on day 5 after treatment. B) FPKM values for leading edge genes of cell cycle pathway increased in diabetic insulin + Glut2 low cells.

2 Supplementary Figure 2. Expansion of human diabetic PMP population and their increased proliferation capacity is maintained throughout multiple passaging. Frequencies and diameters of clonal spheres produced from individual healthy (n = 1) and diabetic (n = 3) donors in secondary (2 ) and tertiary (3 ) passages. The primary (1 ) spheres produced after 2 week culture of primary islet cells were handpicked, pooled, dissociated and cultured at clonal density (5000 cells/ml) to form secondary spheres (A). The same procedure was repeated on 2 spheres after two weeks to produce 3 spheres (B). The individual data from diabetic donors were combined and compared with the single non-diabetic control. Two way ANOVA showed a significant interaction of diabetes and sphere diameter in both passages (F 2 (2,231) = 20.35, p<0.0001, and F 3 (2,108) = 28.21, p<0.001) with a main effect of diabetes on the total number of spheres(f 2 (1,231) = 44.63, p<0.0001, and F 3 (1,108) = 22.07, p<0.001). N = observed wells/group and values indicate means +/- SEMs.

3 Supplementary Figure 3. The proportion of hormone + cells produced from human PMP single colonies increases in diabetic condition. Single clonal spheres produced from three type 1 diabetic (D) and two control (C) human pancreatic islets were differentiated in serum containing media for 2 weeks and were stained for insulin (A) or glucagon (B) expression. (A) Clonal spheres from diabetic samples produced greater numbers of insulin + cells (One way ANOVA, F (4,53) = 5.024, p = 0017). Tukey post test revealed a significant increase of insulin + cell production by D2 and D3 spheres (p<0.05). (B) Spherical colonies from D1 and D3 produced significantly higher proportion of glucagon + cells (One way ANOVA, F (4,68) = 9.701, p<0.0001, Tukey post-test, p<0.05). Number of analyzed spherical colonies is per sample in both experiments. Values indicate means+/-sems.

4 Supplementary Figure 4. PMP population expands in aged human islets. Dissociated pancreatic islet cells from 4 different donors (43, 45, 65, and 68 years old) were cultured at clonal density to form clonal spheres. The frequencies and diameters of spherical colonies were assessed for each sample. Values indicate means+/-sems

5 Supplementary Table 1. Log2 FDR values for comparing the KEGG and REACTOME defined biological pathways of insulin + Glut2 low versus insulin + Glut2 high cells from non-diabetic MIP-GFP transgenic islets at p<0.05 Pathway Glut2 low vs Glut2 high (CTRL) REACTOME Neuronal System KEGG Oxidative Phosphorylation REACTOME Respiratory Electron Transport ATP Synthesis by Chemiosmotic Coupling and Heat Production by Uncoupling Proteins REACTOME Metabolism Of Proteins REACTOME Transmission Across Chemical Synapses KEGG Parkinsons Disease REACTOME Respiratory Electron Transport REACTOME Regulation Of Gene Expression In Beta Cells KEGG Maturity Onset Diabetes Of The Young KEGG Huntingtons Disease KEGG Protein Export REACTOME Regulation Of Insulin Secretion REACTOME Tca Cycle and Respiratory Electron Transport REACTOME Synthesis Secretion and Inactivation Of GIP REACTOME Diabetes Pathways REACTOME Asparagine N Linked Glycosylation REACTOME Regulation Of Beta Cell Development REACTOME Integration Of Energy Metabolism KEGG_Alzheimers Disease REACTOME Unfolded Protein Response REACTOME Neurotransmitter Release Cycle REACTOME Glucose Metabolism REACTOME Gaba Synthesis Release Reuptake and Degradation REACTOME N Glycan Trimming In The ER and Calnexin Calreticulin Cycle

6 REACTOME Incretin Synthesis Secretion and Inactivation REACTOME Post Translational Protein Modification REACTOME Activation Of Chaperone Genes by Xbp1s REACTOME Calnexin Calreticulin Cycle REACTOME Membrane Trafficking REACTOME Synthesis Secretion and Inactivation Of GLP REACTOME SRP Dependent Co-translational Protein Targeting To Membrane KEGG Cardiac Muscle Contraction KEGG Valine Leucine and Isoleucine Degradation REACTOME Neurotransmitter Receptor Binding and Downstream Transmission In The Postsynaptic Cell REACTOME Cytosolic Sulfonation Of Small Molecules REACTOME Translation REACTOME Glutamate Neurotransmitter Release Cycle REACTOME Il-35 and GM-CSF Signaling KEGG Axon Guidance REACTOME Yap1 and Wwtr1 Taz Stimulated Gene Expression REACTOME PPARA Activates Gene Expression REACTOME Hyaluronan Metabolism REACTOME Cell Extracellular Matrix Interactions REACTOME Class A Rhodopsin Like Receptors REACTOME Peptide Ligand Binding Receptors REACTOME Antigen Activates B-Cell Receptor Leading To Generation Of Second Messengers REACTOME Initial Triggering Of Complement REACTOME Immunoregulatory Interactions Between A Lymphoid and A Non Lymphoid Cell KEGG Nod Like Receptor Signaling Pathway

7 REACTOME GPVI Mediated Activation Cascade REACTOME Toll Receptor Cascades REACTOME Transcriptional Regulation Of White Adipocyte Differentiation REACTOME Rig I Mda5 Mediated Induction Of IFN- Alpha Beta Pathways REACTOME Glycosaminoglycan Metabolism REACTOME TRAF6 Mediated IRF7 Activation KEGG T Cell Receptor Signaling Pathway KEGG Natural Killer Cell Mediated Cytotoxicity REACTOME Signaling by Rho GTPases KEGG Rig I Like Receptor Signaling Pathway KEGG Leukocyte Transendothelial Migration REACTOME Chemokine Receptors Bind Chemokines REACTOME Platelet Activation Signaling and Aggregation KEGG TGF Beta Signaling Pathway KEGG Cytosolic DNA Sensing Pathway REACTOME Semaphorin Interactions KEGG Autoimmune Thyroid Disease KEGG Primary Immunodeficiency KEGG JAK-STAT Signaling Pathway KEGG Fc Epsilon RI Signaling Pathway KEGG Chemokine Signaling Pathway REACTOME Developmental Biology REACTOME Axon Guidance REACTOME Cell Surface Interactions At The Vascular Wall KEGG Asthma REACTOME Integrin Cell Surface Interactions REACTOME Signaling by ILS KEGG Fc Gamma R Mediated Phagocytosis

8 KEGG Allograft Rejection REACTOME Interferon Alpha Beta Signaling KEGG Regulation Of Actin Cytoskeleton REACTOME Collagen Formation KEGG Graft Versus Host Disease REACTOME Interferon Signaling KEGG B-Cell Receptor Signaling Pathway REACTOME Interferon Gamma Signaling KEGG Pathways In Cancer KEGG ECM Receptor Interaction KEGG Complement and Coagulation Cascades REACTOME Immune System REACTOME Extracellular Matrix Organization KEGG Focal Adhesion KEGG Toll Like Receptor Signaling Pathway REACTOME Innate Immune System REACTOME Hemostasis KEGG Intestinal Immune Network For IgA Production KEGG Leishmania Infection REACTOME Cytokine Signaling In Immune System KEGG Hematopoietic Cell Lineage KEGG Cytokine Cytokine Receptor Interaction

9 Supplementary Table 2. KEGG and REACTOME defined pathways with significant alteration in insulin + glut2 low cells of STZ-induced diabetic islets. Pathway Glut2 low vs. Glut2 high Glut2 low vs. Glut2 high (Ctrl) (STZ) REACTOME Potassium Channels NA REACTOME Interaction Between L1 and Ankyrins NA REACTOME Amine Derived Hormones NA REACTOME Amine Ligand Binding Receptors NA REACTOME Cell Cycle NA KEGG P53 Signaling Pathway NA REACTOME Inflammasomes NA REACTOME NCAM1 Interactions NA Values are log2 FDR at p<0.05