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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. 6 — a HEK-293F cells were co-transfected with expression vectors harboring Flag-MBP-tagged hMATER (or a blank vector for negative control), Strep-SUMO-tagged hTLE6, Strep-tagged hFLOPED, and HA-tagged 14-3-3γ. 60 h after transfection, cell lysates before (Input) or after immunoprecipitation with anti-Flag affinity agarose gel were immunoblotted for HA-tagged 14-3-3γ, Flag-MBP-tagged hMATER, Strep-SUMO-tagged hTLE6, and Strep-tagged hFLOPED. hMATER, human MATER. hTLE6, human TLE6. hFLOPED, human FLOPED. b Human zygotes were fixed, permeabilized, and incubated with antibodies to TLE6 and pan 14-3-3. The zygotes were imaged by confocal and differential interference contrast (DIC) microscopy. Scale bar, 10 μm. Colocalization of hTLE6 and pan 14-3-3 was presented in the merge. c The in situ interaction between hTLE6 and pan 14-3-3 in human zygotes and 8-cell-stage embryo was determined by proximity ligation assay (PLA) using antibodies to TLE6 and pan 14-3-3 or IgG for isotype control. Scale bar, 10 μm. d HEK-293F cells were co-transfected with expression vectors harboring Flag-MBP-tagged hMATER (or a blank vector for negative control), Strep-SUMO-tagged hTLE6, Strep-tagged hFLOPED, HA-tagged hCDC25B, and either HA-tagged 14-3-3γ or blank vector as denoted. 60 h after transfection, cell lysates before (Input) or after immunoprecipitation with anti-Flag affinity agarose gel were immunoblotted for HA-tagged hCDC25B, HA-tagged 14-3-3γ, Flag-MBP-tagged hMATER, Strep-SUMO-tagged hTLE6, and Strep-tagged hFLOPED. hCDC25B, human CDC25B. e Human zygotes were fixed, permeabilized, and incubated with antibodies to TLE6 and CDC25B. The zygotes were imaged by confocal and DIC microscopy. Scale bar, 10 μm. Colocalization of hTLE6 and hCDC25B was presented in the merge. f The in situ interaction between hTLE6 and hCDC25B in human GV oocytes and embryos was determined by PLA using antibodies to TLE6 and CDC25B or IgG for isotype control. Scale bar, 10 μm.