1. Prevention of Irradiation-induced Salivary Hypofunction by Microvessel Protection in Mouse Salivary Glands 
Ana P Cotrim, Anastasia Sowers, James B Mitchell, Bruce J Baum 
Molecular Therapy 
Volume 15, Issue 12, Pages (December 2007)
DOI: /sj.mt
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Detection of endothelial cells in mouse submandibular gland with and without irradiation (IR). (a) Immunohistochemical detection of endothelial cells. CD31+ (BD Pharmingen) endothelial cells (positive cells stained in blue) are seen in sections of a submandibular gland from untreated mice. (b) Microvessel density (MVD) based on CD31+ cells. The average number of positive cells in 40 fields per gland in the non-irradiated control group and in glands from a group of mice 4 hours after IR is shown. (n = 6 mice/group). CD31+ MVD in salivary glands with 95% confidence intervals (CIs) are shown in a box plot. The upper and lower boundaries of the box represent the 75th and 25th percentile, respectively, of the number of microvessels per field per mouse. The horizontal line within the box represents the median value, and the error bars represent the 95% CIs. Asterisks above and below the bars represent out-of-range values. A total of 12 glands from six mice in each treatment group were studied, and 40 fields were counted/gland. (c) Fluorescence-activated cell sorting (FACS) analyses of non-IR cells and cells (HUVECs) irradiated at 2, 4, and 8 Gy either unstained (u) or stained with CD31 antibody to evaluate if CD31 expression is affected by IR. The bar graph shows the mean ± SEM for the results of FACS analyses with HUVECs. Representative FACS analyses are shown to the right side of panel c. HUVECs, human umbilical vein endothelial cells.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Vector-induced basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) expression in vitro. Data are mean ± SEM of results from three experiments. Enzyme-linked immunosorbent assays were performed with media collected from 293 or A5 cells either not treated, used as control, or cells 48 hours after transduction with either AdbFGF or AdVEGF. Both bFGF and VEGF were undetectable in control cells (not transduced with vectors). Ad, adenovirus.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Vector-induced basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) expression in vivo. Growth factor production was measured by enzyme-linked immunosorbent assay in aqueous extracts of salivary glands obtained 48 hours after delivery of either AdbFGF or AdVEGF as described in Material and Methods. Ad, adenovirus.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Microvessel density in murine salivary glands with and without vector treatment. In these experiments microvascular endothelial cells were detected by measuring aquaporin-1 positive cells. Microvessel density in salivary glands with 95% confidence intervals (CIs) is shown in a box plot. The upper boundary of the box represents the 75th percentile of the number of microvessels per field per mouse. The lower boundary of the box represents the 25th percentile of the data distribution. The horizontal line within the box represents the median value, and the error bars represent the 95% CIs. The asterisks above and below the bars represent out-of-range values. A total of eight glands from four mice in each treatment group were studied, and 40 fields were counted/gland. See Materials and Methods for details. Ad, adenovirus.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
Effect of serotype 5 adenoviral (Ad5) vector administration before irradiation (IR) on salivary flow 8 weeks after IR. AdLacZ, an Ad5 vector–encoding Escherichia coli β-galactosidase; AdbFGF, Ad5 vector–encoding basic fibroblast growth factor (bFGF); AdVEGF, Ad5 vector–encoding vascular endothelial growth factor (VEGF). Mice (n = 10) received either no vector or 5 × 109 particles of the indicated adenoviral vector in each submandibular gland 48 hours prior to IR. After 8 weeks, salivary flow was measured as described in Materials and Methods.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions