Down-regulation of DJ-1 protein in the ejaculated spermatozoa from Chinese asthenozoospermia patients Chun-Na An, M.S., Hui Jiang, M.D., Qi Wang, M.S.,

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Down-regulation of DJ-1 protein in the ejaculated spermatozoa from Chinese asthenozoospermia patients Chun-Na An, M.S., Hui Jiang, M.D., Qi Wang, M.S., Ren-Pei Yuan, M.B., Jian-Ming Liu, M.M., Wan-Lu Shi, B.S., Zi-Yu Zhang, B.S., Xiao-Ping Pu, Ph.D. Fertility and Sterility Volume 96, Issue 1, Pages 19-23.e2 (July 2011) DOI: 10.1016/j.fertnstert.2011.04.048 Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Western blotting analysis of sperm DJ-1 from control subjects (n = 9) and Chinese asthenozoospermia patients (n = 30). (A) DJ-1 expression detected by Western blotting. (B) Levels of DJ-1 were normalized against GAPDH and expressed as fold change relative to control subjects. ∗P<.05 by Student unpaired t test. Fertility and Sterility 2011 96, 19-23.e2DOI: (10.1016/j.fertnstert.2011.04.048) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Quantitive ELISA analysis of DJ-1 levels in ejaculated spermatozoa. (A) Sperm DJ-1 concentration in control subjects (n = 58) and asthenozoospermia patients (n = 113). ∗P<.05 by Student unpaired t test. (B) Sperm DJ-1 levels in control subjects (n = 58), mild asthenozoospermia patients (n = 70), and moderate asthenozoospermia patients (n = 43). ∗P<.05; ∗∗P<.01; by analysis of variance with post hoc Bonferroni test. Fertility and Sterility 2011 96, 19-23.e2DOI: (10.1016/j.fertnstert.2011.04.048) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Dectection of DJ-1 isoelectric point (pI). (A) Representative result of immunoblotting analysis. (B) Proportion of DJ-1 with different pIs. Data were representative of three independent experiments. ∗P<.05 versus control subjects by Student t test. Fertility and Sterility 2011 96, 19-23.e2DOI: (10.1016/j.fertnstert.2011.04.048) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Superoxide dismutase (SOD) activity in spermatozoa. (A) Sperm SOD activity in control subjects (n = 58) and asthenozoospermia patients (n = 113). ∗∗P<.01 by Student unpaired t test. (B) Sperm SOD activity in control subjects (n = 58), mild asthenozoospermia patients (n = 70), and moderate asthenozoospermia patients (n = 43). ∗P<.05; ∗∗P<.01; by analysis of variance with post hoc Bonferroni test. Fertility and Sterility 2011 96, 19-23.e2DOI: (10.1016/j.fertnstert.2011.04.048) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions