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Cell membrane proteins from oviductal epithelial cell line protect human spermatozoa from oxidative damage Venus W. Huang, B.Sc., Weie Zhao, M.B.B.S., Cheuk-Lun Lee, Ph.D., Cherie Y.L. Lee, Ph.D., Kevin K.W. Lam, Ph.D., Jennifer K.Y. Ko, M.B.B.S., William S.B. Yeung, Ph.D., Pak-Chung Ho, M.D., Philip C.N. Chiu, Ph.D. Fertility and Sterility Volume 99, Issue 5, Pages e3 (April 2013) DOI: /j.fertnstert Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions
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Figure 1 Binding of Alexa Fluor (AF) 488–conjugated OE-E6/E7 membrane proteins to human spermatozoa. Uncapacitated or capacitated sperm were incubated with 100 μg/mL AF-488–labeled OE-E6/E7 membrane proteins for 1 hour, followed by the flow cytometric analysis. Representative flow cytometric plots of five replicate experiments are shown. Data are mean ± SEM. *P<.05 compared with the uncapacitated sperm. Fertility and Sterility , e3DOI: ( /j.fertnstert ) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions
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Figure 2 (A) Effects of 100 μg/mL OE-E6/E7 membrane proteins on lipid peroxidation induced by 180 μmol/L H2O2. Lipid peroxidation is presented as percentage changes relative to the control without treatment. (B) Effects of 100 μg/mL OE-E6/E7 membrane proteins on 180 μmol/L H2O2-induced DNA fragmentation. Comet analysis was used to evaluate DNA fragmentation which is presented as percentage tail DNA and tail moment. Data represent the mean ± SEM of five separate experiments using five different sperm samples. *P<.05 compared with the control without treatment. #P<.05 compared with the H2O2-alone treatment group without OE-E6/E7 membrane proteins. Fertility and Sterility , e3DOI: ( /j.fertnstert ) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions
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Figure 3 (A) Effects of 100 μg/mL OE-E6/E7 membrane proteins on 180 μmol/L H2O2–induced intracellular ROS level. Intracellular reactive oxygen species (ROS) level is presented as percentage change relative to the control. (B) Effects of 100 μg/mL OE-E6/E7 membrane proteins on sperm superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activiy. Antioxidant activity is presented as percentage change relative to the control without treatment. Data represent the mean ± SEM of five separate experiments using five different sperm samples. *P<.05 compared with the control without treatment. #P<.05 compared with the H2O2-alone treatment group without OE-E6/E7 membrane proteins. Fertility and Sterility , e3DOI: ( /j.fertnstert ) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions
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Supplemental Figure 1 Content of glycogen synthase kinase 3 beta (GSK-3β) and E-cadherin in total cell lysate, cytosolic protein fraction, and membrane protein fraction of OE-E6/E7 cells. Top: Representative Western blot of three replicate experiments. Bottom: Semiquantification of the protein bands by densitometry. Data represent mean ± SEM of three separate experiments. Fertility and Sterility , e3DOI: ( /j.fertnstert ) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions
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