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. 2021 Feb 11;8:598634. doi: 10.3389/fcell.2020.598634

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Copyright © 2021 Ranawakage, Okada, Sugio, Kawaguchi, Kuninobu-Bonkohara, Takada and Kamachi.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Expression of the tagged Sox3 protein. (A) Western blot and HiBiT blot analyses of the tagged Sox3 protein. Two-color Western blot analysis of sox3 knock-in embryos derived from the FLAGx3-50_#16 (a) and PAx3-50_#21 founders (b) with antibodies against Sox3, FLAG tag or PA tag, and α-tubulin. HiBiT blotting using an excessive amount of LgBiT protein and substrate to detect the C-terminal HiBiT peptide of the tagged Sox3 proteins (c). (B) Whole-mount immunohistochemical staining for the tagged Sox3 protein. Each primary antibody used to stain zebrafish is indicated below each image. Genotyping was conducted using PCR with genomic DNA prepared from the immunostained embryos after image acquisition.