1. Toward gene therapy of endometriosis: adenovirus-mediated delivery of dominant negative estrogen receptor genes inhibits cell proliferation, reduces cytokine production, and induces apoptosis of endometriotic cells 
Essam-Eldin R. Othman, M.D., Salama Salama, Ph.D., Nahed Ismail, Ph.D., M.D., Ayman Al-Hendy, Ph.D., M.D. 
Fertility and Sterility 
Volume 88, Issue 2, Pages (August 2007)
DOI: /j.fertnstert
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

2. FIGURE 1
Coxsackievirus-adenovirus receptor mRNA expression by endometriosis and endometrial cells. (A): Reverse transcription-polymerase chain reaction amplification of CAR mRNA expression by normal endometrial cells (lanes 1 to 5) or endometriotic cells (lanes 6 to 10). Using the same RNA template concentration (250 μg/μL), endometriotic cells expressed higher levels of the CAR mRNA as compared with normal endometrial cells. The expressed band is at the expected size of 342 bp in both cell types. (B): Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA expression by endometrial and endometriotic cells as internal control.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

3. FIGURE 2
(A): Endometriotic cells. (B): Endometrial cells. Transfectability of human endometriotic and endometrial cells by adenovirus. Endometriotic and endometrial cells are transfected with Ad-Lac Z at a multiplicity of infection of 50 PFU/ cell followed by staining with X-gal. Nearly all of the endometriotic cells showed positive blue nuclear staining. The endometrial cells were highly confluent, with only a few cells showing the positive staining.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

4. FIGURE 3
Morphology of endometriotic cells after transfection with different types of viruses. (A): Endometriotic cells 3 days after transfection with Ad-DN-ER at an MOI of 50 PFU/cell. The cells show rounding, irregular outlines, and cellular detachment. (B): Endometriotic cells 3 days after transfection with Ad-Lac Z at an MOI of 50 PFU/cell. (C): Control endometriotic cells not transfected with any virus. Cells in B and C are healthy and actively growing with no visible pathological changes.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

5. FIGURE 4
Cell proliferation assay of endometriotic cells treated by different viruses. Endometriotic cells treated with Ad-DN-ER, at an MOI of 50 PFU/cell, showed an initial brief increase in cell numbers followed by a progressive decrease in cell numbers over time. Cells treated with Ad-Lac Z at the same MOI and untreated cells continued to proliferate exponentially. *P<.05 compared with controls.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

6. FIGURE 5
Effect of Ad-DN-ER on cytokine production by endometriotic cells. Significantly less MCP-1, VEGF, and IL-6 quantities were secreted by Ad-DN-ER–treated endometriosis cells than Ad-Lac Z–treated cells. Both viruses were used at an MOI of 50 PFU/cell. Measurements were conducted on cell culture media samples collected on the third day after virus transfection. *P<.05 compared with controls.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

7. FIGURE 6
A TUNEL assay to test the effect of Ad-DN-ER on induction of apoptosis in endometriotic cells. (A): Ad-DN-ER–treated cells. Most of the cells are positive for apoptosis (brown nuclei). (B): Ad-Lac Z–treated cells (both viruses were used at a multiplicity of infection of 50 PFU/cell). (C): Cells treated with medium only (no virus added). Most cells in B and C are negative for apoptosis (blue nuclei) and are stained with hematoxylin counterstain.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

8. FIGURE 7
Effect of Ad-DN-ER on the induction of apoptosis in human endometriotic cells. The Ad-DN-ER–treated endometriosis cells (at an MOI of 50 PFU/cell) showed a higher percentage of apoptotic cells than the two control groups as detected by TUNEL assay. *P<.05.
Othman. Gene therapy of endometriosis. Fertil Steril 2007.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions