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. 2017 Aug 21;7:8874. doi: 10.1038/s41598-017-09262-6

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© The Author(s) 2017

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/.

PMC Copyright notice

Conserved cysteines in Prdx1 are essential for pronephros development. (A) Three prdx1 mutants, namely, C53S (M1), C173S (M2), and C53S/C173S (M1/2), were subcloned using PCR-based site-directed mutagenesis. (B) Flag-tagged Prdx1* or HA-tagged prdx1 (200 pg) was co-injected into both blastomeres of two-cell stage embryos. Lysates from stage 12 embryos were used for immunoprecipitation. WT Prdx1 only showed in immune complex whereas mutant Prdx1 did not. (C) prdx1 MOs (40 ng) were co-injected with WT or mutant prdx1 (M1, M2 or M1/M2) into both blastomeres of two-cell stage embryos. Embryos at the stage 33 were used for whole-mount in situ hybridization, and proximal tubules in the pronephros were visualized with a smp30 probe. Inhibited proximal tubule formation in prdx1 morphants was rescued by WT prdx1 but not with mutant prdx1* RNAs. (D) Graphical demonstration of smp30 expression in embryos co-injected with WT or mutant prdx1 (M1, M2 or M1/M2) and prdx1 MOs compared with the controls.