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. 2010 Mar 18;285(20):15627–15636. doi: 10.1074/jbc.M109.079483

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Inhibitory effect of dicalcin on in vitro fertilization. A, effect of pretreatment with anti-dicalcin antibody on fertilization. Ovulated eggs were pretreated with preimmune antibody (Pre, 50 mg/liter) or anti-dicalcin (Anti-dica, 5 and 50 mg/liter) followed by incubation of sperm. The number of two-cell embryos was counted until one of the embryos proceeded to the four-cell-stage. Fertilization success was scored and normalized (n = 6; *, p = 0.007). B, effect of pretreatment with dicalcin on fertilization. Ovulated eggs were pretreated with BSA or dicalcin at indicated concentrations followed by insemination (n = 6; *, p = 0.004; **, p = 7.2 × 10⁻⁵). EGTA: addition of EGTA (16 μm) prior to inseminations. +Anti-dica: addition of anti-dicalcin antibody (50 mg/liter) together with dicalcin. C, effect of preincubation of sperm with dicalcin on fertilization. Ovulated eggs were inseminated with sperm (SP) that were preincubated with dicalcin (Dica). As a control, eggs were pretreated with BSA (BSA) similarly as described above and inseminated with sperm.