1. Volume 9, Issue 2, Pages 198-208 (February 2004)
Development of Efficient Viral Vectors Selective for Vascular Smooth Muscle Cells 
Lorraine M Work, Stuart A Nicklin, Nick J.R Brain, Kate L Dishart, Dan J Von Seggern, Michael Hallek, Hildegard Büning, Andrew H Baker 
Molecular Therapy 
Volume 9, Issue 2, Pages (February 2004)
DOI: /j.ymthe
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

2. Fig. 1
Phage display biopanning recovery studies and sequence analyses. (A) In vitro biopanning protocol. Briefly, phage were incubated with target cells, eluted, amplified, and titered (all rounds except round 2). Round 2, preclearing, was performed as shown (2). Peptide inserts were sequenced after round 4 for linear libraries and 3 for cyclic. (B) Phage recoveries following each successive round of biopanning with either the linear or the constrained library (2 × 1011 pfu phage input at each round) were determined from triplicate wells. *P < 0.05 vs the previous round of biopanning. (C) Comparison of peptide sequences isolated from HSVSMC and HSVEC. Numbers shown are peptide inserts sequenced, with the number of identical sequences shown in parentheses. Numbers in crossover indicate common peptides.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

3. Fig. 2
Assessment of Ad vectors and transduction in human SMC. HSVSMC were exposed (A) for 3 h to increasing doses of peptide-modified Ads or (B) for short (10–60 min) exposure times before quantification 72 h postinfection. HCASMC were exposed (C) for 3 h to increasing doses of peptide-modified Ads or (D) for short (10–60 min) exposure times. *P < 0.05 vs RAd CTL and #P < 0.05 vs RAd KO1 at the same dose or time point.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

4. Fig. 3
RAd EYH transduction is maintained cross species, independent of CAR and selective against HSVEC. (A) Porcine SVSMC were exposed to 20,000 particles/cell of peptide-modified or control Ad for the indicated times. (B) HSVSMC were infected with indicated virus in the presence of an anti-CAR antibody or nonimmune serum control. (C) HSVEC were exposed to 50,000 particles/cell of peptide-modified Ads for short (10–60 min) exposure times. Reporter gene expression was determined fluorimetrically 72 h postinfection. *P < 0.05 vs RAd CTL and #P < 0.05 vs RAd KO1 at the same time point.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

5. Fig. 4
Assessment of AAV transduction in human SMC. (A) HSVSMC or (B) HCASMC were exposed to 20,000 genome particles/cell of peptide-modified AAV for the indicated times. Reporter gene expression was determined fluorimetrically 72 h postinfection. (C) HSVSMC were exposed to AAV CTL or AAV EYH at 4°C for 1 h and virion binding was quantified by real-time PCR. *P < 0.05 vs AAV CTL at the same time point.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

6. Fig. 5
HSPG dependence and cell selectivity of AAV EYH. (A) Heparin-affinity column analysis. Top: Lane 1, positive control; lane 2, negative control; lane 3, AAV CTL; lane 4, flowthrough; lanes 5–7, sequential washing steps; lanes 8 and 9, sequential elutions with 1 M NaCl. Bottom: Same as the top except lanes 1 and 2 are empty and lane 3 contains AAV EYH. (B) HSVSMC were infected with AAV CTL or AAV EYH in the absence or presence of IU heparin/1 × 106 virions for 18 h and analyzed for reporter gene expression 72 h later. (C) HSVEC transduction by peptide-modified AAVs for the indicated times. Transgene expression was determined fluorimetrically 72 h postinfection. *P < 0.05 vs transduction in the absence of heparin or vs AAV CTL. (D) Tropism modification by AAV EYH and AAV SIG [11] in HSV EC and SMC compared to AAV CTL. *Significantly higher than AAV CTL and **significantly lower than AAV CTL.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions

7. Fig. 6
Sensitivity of AAV CTL and AAV EYH to proteasome inhibitors. HCASMC were exposed to 20,000 gp/cell AAV CTL or AAV EYH in the absence or presence of 40 μM LLnL or 40 μM mg132 and transgene expression was quantified at 24 h. *Statistically different from transduction in the absence of inhibitor for each AAV.
Molecular Therapy 2004 9, DOI: ( /j.ymthe )
Copyright © 2003 American Society of Gene Therapy Terms and Conditions