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. 2019 Oct 1;10:4456. doi: 10.1038/s41467-019-12406-z

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© The Author(s) 2019

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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Fig. 2 — BAI1/3 and CD36 are expressed on oocytes and contribute to fertilization. a Schematic for the assay to determine PtdSer recognition moieties on mouse oocytes. b Oocytes-bound multiple untreated carboxylate beads within the microvillar region (left), but not the BAI1-TSR-treated beads (right). Scale bar, 20 μm. c Summary of three experiments using carboxylate binding oocytes. ***p < 0.001 (Two-tailed unpaired Student’s t-test). d, PtdSer expression (qPCR) on cumulus-free Metaphase II oocytes from wild-type female mice (n = 4 experiments). e Concanavalin A (ConA) staining (marking microvilli) overlap with staining for BAI1/3 and CD36 on ZP-free oocytes. Scale bar, 20 μm. f Immunofluorescence (top) and immunohistochemistry (bottom) staining of mouse ovaries with BAI1/3 antibodies. Connexin-43 (green) marks follicular cells, ovarian follicles (dotted lines) are indicated. Arrows: oocytes in secondary or tertiary follicles; arrowheads: oocytes in primary follicles. Scale bar, 50 μm. g, Schematic of fertilization assays assessing the role of BAI1/3 and CD36. h, i Multiple two-cell embryos are observed in the control group (left, arrows) but reduced after CD36 and BAI1/3 antibody treatment (right). Scale bar, 100 μm. The compilation of data from 7 independent experiments is shown in i. Each dot represents one experiment, with matching experiments shown (shape and color). sIgG, sheep IgG, mIgA, mouse IgA. **p < 0.05 (two-tailed unpaired Student’s t test). j Schematic of in vitro fertilization assays to test BAI1/3 and CD36 in sperm entry. Untreated or antibody-treated oocytes were loaded with DAPI, and mixed with sperm. The percentage of oocytes with decondensed sperm DNA was evaluated after 3 h. k, l A representative image of a fertilized oocyte with one decondensed sperm DNA k. Scale bar: 20 μm. Quantitation of oocytes with decondensed sperm DNA after blocking with CD36 and BAI1/3 antibodies l. Each dot represents one experiment (n = 3 independent experiments) and the matching experiments are shown (shape and color). **p < 0.01 (two-tailed unpaired Student t test). Data are presented as mean ± s.e.m. Source Data are provided in the Source Data File