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. 2020 Mar 24;10:5335. doi: 10.1038/s41598-020-62091-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Anti-hSPACA6 antibody inhibited Human fertilization. Overnight (18 h) insemination of zona-free human oocytes with human sperm in the presence of rabbit polyclonal anti-hSPACA6 (10 µg/ml), or rabbit IgG (10 µg/ml) or only IVF medium. The number of fused sperm heads into egg cytoplasm was recorded after insemination under fluorescent microscope (Nikon eclipse E600) and the fertilization index (FI: Number of decondensed sperm per oocyte, mean ± SEM) was calculated. FI decreased drastically from 29.5 ± 2.5 (n = 11) for the control group to 10.1 ± 1.1 (n = 11) for antibody treated group (10 µg/ml). No significant difference was observed between IVF medium control and IgG (at 10 µg/ml) control groups (FI: 26.4 ± 5.5 (n = 7)). Examples of illustrative images obtained after confocal analysis (Spinning Disk) are shown for the control and antibody groups. Transmission images were superimposed with Hoechst fluorescent signals (blue).