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. 2019 Dec 11;9:18845. doi: 10.1038/s41598-019-55449-4

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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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Conjugates between PTD of TCTP and siRNAs are effectively delivered into mouse oocytes. (A) Synthesis of conjugates between PTD of TCTP and siRNAs via a thiol-maleimide coupling. (B) After conjugation between PTD and siRNA, the transparency of reaction mixtures was shown. (C) The conjugation between PTD and siRNA was determined by Ellman’s test. (D) The uptake of PTD-siRNA conjugates was determined by confocal microscopy after treating GV oocytes with PTD-siRNA for 4 hours. Bar, 20 μm. (E) GV oocytes were treated with either unconjugated cyclin B1 siRNAs or PTD-cyclin B1 siRNAs for 24 h with IBMX. Knockdown of cyclin B1 was confirmed by immunoblotting analysis. β-actin was used as a loading control. Each lane contains 40 oocytes. Quantification of cyclin B1 levels was shown. Cropped blots are represented and full-length blots are reported in Supplementary Fig. 2. (F) After 24 hours treatment with either unconjugated cyclin B1 siRNAs or PTD-cyclin B1 siRNAs, oocytes were released from IBMX-mediated GV arrest and GV breakdown (GVBD) was scored after 4 hours. Representative images from three independent experiments are shown, along with quantification of GVBD rate. Bar, 80 μm. The data are expressed as mean ± SEM from three independent experiments. *p < 0.05, **p < 0.001 (Student’s t-test).