1. Volume 2, Issue 4, Pages 381-386 (October 2000)
Downregulation of CXCR4 Gene Expression in Primary Human Endothelial Cells Following Infection with E1−E4+ Adenovirus Gene Transfer Vectors 
Ramachandran Ramalingam, Stefan Worgall, Shahin Rafii, Ronald G. Crystal 
Molecular Therapy 
Volume 2, Issue 4, Pages (October 2000)
DOI: /mthe
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

2. FIG. 1
Downregulation of CXCR4 expression in HUVEC following infection with an E1–E4+ Ad vector. (A) Northern analysis. Total RNA (10 μg/lane) was isolated from control uninfected cells and cells infected with E1–E4+ Ad vector (48 h). The blot was then hybridized to a CXCR4 probe. Hybridization with a control GAPDH probe demonstrated integrity of RNA and equal loading. (B) Flow cytometry analysis of CXCR4 expression on the endothelial cell surface. Control uninfected cells and cells infected with E1–E4+ Ad were treated with either a control IgG or PE-labeled antibody against CXCR4 and then analyzed by flo
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

3. FIG. 2
Flow cytometry analysis demonstrating E1–E4+ Ad vector-mediated downregulation of CXCR4 in endothelial cells compared to dendritic cells (DC) and Bcl3 myeloma cells. Control uninfected cells and cells infected with E1–E4+ Ad vector were treated with PE-labeled antibody against CXCR4 and analyzed by flo
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

4. FIG. 3
Lack of downregulation of CXCR4 expression on endothelial cells following infection with E1–E4– Ad. (A) Northern analysis of CXCR4 gene expression in endothelial cells. Total RNA (10 μg/lane) was isolated from uninfected control cells and cells infected with E1–E4+ Ad or E1–E4– Ad vector (48 h). The blot was then hybridized to CXCR4 and control GAPDH probes. Densitometry of CXCR4/GAPDH was 0.61 RLU for lane 1, 0.18 for lane 2, and 0.61 for lane 3, respectively. (B) Flow cytometry analysis of CXCR4 expression on the endothelial cell surface. Control uninfected cells and cells infected with E1–E4+ Ad or E1–E4– Ad were treated with either control IgG or PE-labeled antibody against CXCR4 and then analyzed by flo
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

5. FIG. 4
Time course of downregulation of CXCR4 expression in endothelial cells. HUVECs were infected with E1–E4+ Ad vector or E1–E4– Ad vector and were harvested for RNA isolation either immediately after 90 min infection (0 h) or 12, 24, or 48 h postinfection. Control cells were exposed to infection medium alone. Total RNA was isolated from both control and Ad vector-infected cells and 10 μg of RNA was used in Northern blot analysis. The blot was then hybridized to either a CXCR4 probe or a control GAPD
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

6. FIG. 5
Chemotaxis of HUVEC to SDF-1α following infection with E1–E4+ Ad or E1–E4– Ad. Migration of uninfected control cells or cells exposed to E1–E4+ Ad or E1–E4– Ad vector toward SDF-1α was quantified by counting triplicate wells and expressed as a mean value compared to input cel
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions