1. Volume 26, Issue 3, Pages 568-575.e3 (September 2017)
Persistence of Pancreatic Insulin mRNA Expression and Proinsulin Protein in Type 1 Diabetes Pancreata 
Clive Wasserfall, Harry S. Nick, Martha Campbell-Thompson, Dawn Beachy, Leena Haataja, Irina Kusmartseva, Amanda Posgai, Maria Beery, Christopher Rhodes, Ezio Bonifacio, Peter Arvan, Mark Atkinson 
Cell Metabolism 
Volume 26, Issue 3, Pages e3 (September 2017)
DOI: /j.cmet
Copyright © 2017 Elsevier Inc. Terms and Conditions

2. Cell Metabolism 2017 26, 568-575.e3DOI: (10.1016/j.cmet.2017.08.013)
Copyright © 2017 Elsevier Inc. Terms and Conditions

3. Figure 1
Proinsulin, Insulin, and Glucagon Expression in Type 1 Diabetes
(A–E) Composite images of pancreas sections stained for proinsulin (green), insulin (red), glucagon (yellow), and DAPI (nuclei, blue) are shown for sections containing islets from a control 24-year-old male Caucasian (nPOD 6131) (A), a 12-year-old male African American with T1D for 1 year (nPOD 6052) (B and C), a 23-year-old female Caucasian with T1D for 7 years (nPOD 6070) (D), and a section containing pancreas exocrine tissue from a 12.5-year-old female Caucasian with T1D for 2 years (nPOD 6371) (E).
(F–T) The individual channels for proinsulin (green; F–J), insulin (red; K–O), and glucagon (yellow; P–T) are also shown.
Insets display: proinsulin- and insulin-positive cells in the exocrine pancreas (A), an insulin-negative, proinsulin-negative, glucagon-positive islet (C), and proinsulin-, insulin-, and glucagon-positive cells (E) in the exocrine pancreas.
Scale bars represent 50 μm. See also Figure S1.
Cell Metabolism  , e3DOI: ( /j.cmet )
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4. Figure 2
Total Protein Extracts and Gene Expression from Human Pancreas Sections
(A–E) ELISAs of acid-ethanol extracts from human pancreas tissue sections for insulin (μInternational Units, μIU) (A), proinsulin (B), C-peptide (C), islet amyloid polypeptide (D), and glucagon (E) (values were normalized against total protein). ND, not detected/examined are noted on each graph. Data are presented as median with interquartile range.
(F–H) Correlation between extracted proinsulin and extracted insulin in controls (F), autoantibody-positive (Ab+) (G), and T1D (H) is shown. For T1D subjects, correlation analysis is reported for all data, as well as a separate analysis of the five subjects with insulin detected in the normal range (Detectable INS Only), with the latter represented by the trend line.
(I–N) Real-time qPCR Cq values for control and T1D pancreata for INS (I), unspliced INS heterogeneous nuclear RNA (hnRNA) (J), INS-IGF2 readthrough mRNA (K), IAPP (L), GCG (M), and SST (N) expression were compared. For INS hnRNA, INS-IGF2, and IAPP, statistical analyses were not performed due to the number of T1D samples with undetectable RNA (ND, not detected/examined). Data are presented as median with interquartile range.
p values are indicated on the figure.
Cell Metabolism  , e3DOI: ( /j.cmet )
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5. Figure 3
Evaluation of Insulin/Glucagon IHC, INS ISH, Prohormone Convertase, and Protease Gene Expression from Human Pancreas
(A–J) Insulin (red) and glucagon (blue) protein was detected by IHC (A–E), and insulin mRNA (pink) was detected by ISH (F–J) in pancreas sections from donors without diabetes (A and F; control; nPOD 6172) and T1D pancreata with 7 (B, C, G, and H; nPOD 6070) and 35 year duration (D, E, I, and J; nPOD 6031). Insets in (D) and (E) illustrate glucagon-only-positive islets. Red arrows indicate single insulin-positive cells located in the exocrine tissue and peri-ductal. Scale bars represent 50 μm.
(K–O) For control (n = 5) and T1D subjects (n = 11), IHC analysis of glucagon and insulin within pancreas sections from control donors, as well as short (0–7 years; n = 5) or long (8-35 years; n = 6) duration T1D, were analyzed for insulin-positive cells per islet (K), cell counts within the islets (L and M), and exocrine tissue (N and O). Data are presented as mean ± SD.
(P–R) qPCR for control versus T1D pancreata PCSK1 (P), PCSK2 (Q), and CPE (R). Data are presented as median with interquartile range.
p values indicated on the figure.
Cell Metabolism  , e3DOI: ( /j.cmet )
Copyright © 2017 Elsevier Inc. Terms and Conditions