1. Volume 5, Issue 2, Pages 154-160 (February 2002)
Transduction of Human and Mouse Pancreatic Islet Cells Using a Bicistronic Recombinant Adeno-associated Viral Vector 
Matthias Kapturczak, Sergei Zolotukhin, Jeff Cross, Antonello Pileggi, R. Damaris Molano, Marda Jorgensen, Barry Byrne, Terence R. Flotte, Tamir Ellis, Luca Inverardi, Camillo Ricordi, Harry Nick, Mark Atkinson, Anupam Agarwal 
Molecular Therapy 
Volume 5, Issue 2, Pages (February 2002)
DOI: /mthe
Copyright © 2002 American Society for Gene Therapy Terms and Conditions

2. FIG. 1
Confocal microscopy images of human islets taken 5 days after infection with rAAV2-RFP-IRES-GFP. The top panel represents mock-transduced (control) islets, the bottom panel rAAV2-RFP-IRES-GFP (UF19)-treated islets. Arrows indicate islet cells expressing both RFP and GFP. The red spots in the lower merge panel are background.
Molecular Therapy 2002 5, DOI: ( /mthe )
Copyright © 2002 American Society for Gene Therapy Terms and Conditions

3. FIG. 2
Confocal microscopy images of mouse islets taken 5 days after infection with rAAV2-RFP-IRES-GFP. The top panel represents mock-transduced (control) islets, the bottom panel rAAV2-RFP-IRES-GFP (UF19)-treated islets. Arrows indicate islet cells expressing both RFP and GFP.
Molecular Therapy 2002 5, DOI: ( /mthe )
Copyright © 2002 American Society for Gene Therapy Terms and Conditions

4. FIG. 3
In vitro and in vivo metabolic function of transduced islets. (A) Glucose stimulated insulin release from control and rAAV2-RFP-IRES-GFP-transduced islets at 12 and 24 hours post transduction. The results are expressed as mean ± SEM (n = 3 in each group). (B) Blood glucose monitoring in diabetic (STZ-induced) C57BL/6 recipients of syngeneic pancreatic islet transplant from a representative experiment (n = 2 in each group). Arrow indicates time of islet graft removal.
Molecular Therapy 2002 5, DOI: ( /mthe )
Copyright © 2002 American Society for Gene Therapy Terms and Conditions

5. FIG. 4
Histological evaluation of transplanted islets. Immunohistochemical staining of transplanted mouse islets using a rabbit anti-RFP (A–C) or GFP (D–F) antibody as described in the methods. (A and D) Mock-transduced islets (control). (B and E) rAAV2-RFP-IRES-GFP-transduced islets. (C and F) rAAV2-RFP-IRES-GFP transduced islets stained using the same protocol as in (B) and (E), but with rabbit IgG as primary antibody instead of the RFP or GFP antibody (IgG control). Black arrows indicate examples of RFP (brown color) and GFP-positive (black color) cells. *The underlying renal cortex. The black horizontal bars represent a magnification of 100 μm. Hematoxylin and eosin staining of mock-transduced (G) and rAAV-RFP-IRES-GFP-transduced islets (H). The insets in (G) and (H) show anti-insulin staining (brown color) of respective islet grafts.
Molecular Therapy 2002 5, DOI: ( /mthe )
Copyright © 2002 American Society for Gene Therapy Terms and Conditions

6. FIG. 5
Functional map of pTR-UF19 rAAV2 vector. The elements of the cassette are as follows: TR is the terminal repeat of AAV2 virus; CBA is the cytomegalovirus (CMV) immediate early enhancer linked to chicken β-actin promoter and is derived from pBacMam-1 (Novagen, Madison, WI); DsRed1 is a red fluorescent protein cDNA [32] and is derived from pDsRed1-N1 (Clontech, Palo Alto, CA); IRES is the internal ribosome entry site of poliovirus type 1 from pSBC-1 [33]; GFP is a “humanized” green fluorescent protein cDNA from pTR-UF5 [34]; WPRE is a woodchuck hepatitis virus post-transcriptional regulatory element derived from pBS-WPRE-B11 [45]. Poly(A) is the bovine growth hormone polyadenylation site from pTR-UF2. The size of the individual components is shown. The actual size of the packaged AAV vector is 4.9 kb.
Molecular Therapy 2002 5, DOI: ( /mthe )
Copyright © 2002 American Society for Gene Therapy Terms and Conditions