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. 2024 Oct 15;15:8887. doi: 10.1038/s41467-024-53277-3

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 3 — a Immunoblotting of 14-3-3 in normal control and Tle6^Null germinal vesicle (GV) oocytes. b Immunofluorescence (IF) staining in GV oocytes isolated from control or Tle6^Null female mice. Scale bar, 10 μm. c Immunoblotting of 14-3-3 in control and Floped^Null GV oocytes. d IF staining in GV oocytes isolated from control or Floped^Null female mice. Scale bar, 10 μm. e Immunoblotting of 14-3-3 in zygotes from control and Tle6^Null female mice. Zygotes were flushed from the oviduct of female mice that underwent superovulation with gonadotrophins and were mated with normal males. f GV oocytes isolated from control or Tle6^Null female mice were cultured to mature eggs and incubated with normal sperms in vitro. After in vitro fertilization (IVF), the resulted zygotes were collected for IF staining. Scale bar, 10 μm. g Immunoblotting of 14-3-3 in zygotes from control and Floped^Null female mice. h GV oocytes isolated from control or Floped^Null female mice were cultured to mature eggs and incubated with normal sperms in vitro. After IVF, the resulted zygotes were collected for IF staining. Scale bar, 10 μm. i Representative images of mouse zygotes cultured for 24, 48, 72, and 96 h after co-microinjection with mCherry-Trim21 mRNA + isotype IgG (control) or mCherry-Trim21 mRNA + anti-pan 14-3-3 antibody. Scale bar, 50 μm. The validation for the loss of pan 14-3-3 was presented in Supplementary Fig. 11. j The proportions of embryos on different stages in panel i were calculated. N = 3 independent experiments. The data are presented as mean ± S.D. (standard deviation). k Representative images of zygotes cultured for 24, 48, 72, and 96 h after microinjection with mRNA solvent (control) or mixed mRNAs of seven 14-3-3 paralogs. Zygotes were flushed from the oviduct of Tle6^Null female mice (mated with normal males) at 11–13 h post human chorionic gonadotrophin injection. Scale bar, 50 μm. l, The average proportions of embryos on different stages in panel k were calculated. N = 2 independent experiments.