Determination of Poly (ADP-ribose) polymerase (PARP) homologues in human ejaculated sperm and its correlation with sperm maturation Rajesh Jha, Ph.D.,

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Determination of Poly (ADP-ribose) polymerase (PARP) homologues in human ejaculated sperm and its correlation with sperm maturation Rajesh Jha, Ph.D., Ashok Agarwal, Ph.D., HCLD., Reda Mahfouz, M.D., Uwe Paasch, M.D., Sonja Grunewald, M.D., Edmund Sabanegh, M.D., Satya P. Yadav, Ph.D., Rakesh Sharma, Ph.D. Fertility and Sterility Volume 91, Issue 3, Pages 782-790 (March 2009) DOI: 10.1016/j.fertnstert.2007.12.079 Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Schematic representation of the experimental study design. Fertility and Sterility 2009 91, 782-790DOI: (10.1016/j.fertnstert.2007.12.079) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Detection of PARP proteins 75 kDa, 63 kDa, and 60 kDa, employing Western blot and their densitometric analysis for evaluating the protein level in donors and infertile men between (A) immunoblot of mature and immature sperm fraction; (B) 75 kDa band intensity in mature and immature sperm fraction; (C) 60 and 63 kDa band intensity in mature and immature sperm fraction; and (D) validation of primary antibody specificity by immunoblotting with secondary antibody alone. Fertility and Sterility 2009 91, 782-790DOI: (10.1016/j.fertnstert.2007.12.079) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Immunolocalization study of PARP-1 on (A) mature and (B) immature donor sperm, and (C) mature and (D) immature sperm from infertile patients. Crossreactivity was tested by incubating with secondary antibody alone (E). Fertility and Sterility 2009 91, 782-790DOI: (10.1016/j.fertnstert.2007.12.079) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Analysis of PARP proteins by Western blotting after induction of apoptosis by H2O2 and staurosporine in (A) mature and (B) immature sperm fraction from donors and infertile patients. Quantitative PARP protein level are represented in graphs (C) ∼75 kDa, (D) ∼63 kDa, and (E) ∼60 kDa. (F) Apoptosis was measured by externalization of phosphatidylserine/Annexin V staining. Fertility and Sterility 2009 91, 782-790DOI: (10.1016/j.fertnstert.2007.12.079) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions