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. 2023 Jun 22;14:3729. doi: 10.1038/s41467-023-39256-0

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© The Author(s) 2023

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. 8 — a Venn diagram showing the overlapping of transcripts that were stabilized during GV-to-MII transition in Cnot6l−/− and Nat10-ZcKO MII oocytes (FC = [WT MII/Nat10-ZcKO MII]≥2, p < 0.05). b Fold change of relative expression levels of transcripts encoding ribosomal protein subunits in Nat10-ZcKO relative to WT oocytes at the MII stage. The values of log2(FC[Nat10-ZcKO/WT]) are listed in the right column. c qPCR results showing the relative levels of indicated transcripts (Cnot6l, Cnot7 and Btg4) in WT and Nat10-ZcKO oocytes at MII stage. Data are presented as the mean ± SEM, n = 3. n.s., non-significant, **p < 0.01 by two-tailed Student’s t-test. Cnot6l, p = 0.0906; Cnot7 p = 0.0020; Btg4 p = 0.0031. d Western blot displaying the NAT10, CNOT6L, CNOT7 and BTG4 protein levels in MII oocytes of WT and Nat10-ZcKO mice. α-TUBULIN was used as a loading control. n = 3 biologically independent samples were included in each group. e A schematic illustration depicting the design strategy and the key steps for Hairpin Adaptor-Poly(A) Tail length (HA-PAT) assay. The 1st strand of cDNA was synthesized with the hairpin adaptor (HA) primer in conjunction with a P5TSO primer containing three “G”, via a mechanism of “template-switching”. GSP, Gene-specific primer; A0, the PCR product resulting from the amplification with a gene-specific pair of GSPxF and GSPxR primers; GSPxR primer was designed against an mRNA’s 3′ terminals preceding the poly(A) sequence; poly(A)-containing PCR products were amplified with GSPxF and fixed HAPrimerR primers. The full sequence for hairpin adaptor (HA) is listed at the bottom. W indicates degenerate nucleotides (A or T); * The asterisk indicates the phosphorothioate modification. f, g HA-PAT assay results showing changes in poly(A)-tail lengths of indicated transcripts in WT and Nat10-ZcKO oocytes at GV and MII stages. Experiments were performed in triplicates; a representative image is shown in the 2% agarose gel in f and the length distribution is shown in the densitometric curves in (g). n = 3 biologically independent samples were included in each group (d, f). Source data are provided as a source data file.