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. 2021 Apr 22;12:2395. doi: 10.1038/s41467-021-22604-3

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© The Author(s) 2021

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. 3 — a Gene copy numbers of different osmoregulation related genes among arthropods^{123, 124}. F-H-ATPase is the only gene family that significantly expands in the E. sinensis genome. The species in the comparison includes E. sinensis (ESIN), Portunus trituberculatus (PTRI), Procambarus virginalis (PVIR), Parhyale hawaiensis (PHAW), Daphnia pulex (DPUL), Drosophila melanogaster (DMEL), Bombyx mori (BMOR), Tribolium castaneum (TCAS), Strigamia maritima (SMAR), Mesobuthus martensii (MMAR), Tetranychus urticae (TURT), Limulus polyphemus (LPOL), and Patinopecten yessoensis (PYES). The osmoregulation related genes include Na⁺/K⁺ ATPase (NKA), Na⁺/K⁺/2Cl^- cotransporter (NKCC), Na⁺/K⁺/Ca²⁺ exchanger (NKCaE), Na⁺/K⁺ symporter (Na/K-sympoter), sodium/glucose cotransporter (Na/glucose), Ca²⁺ transporting ATPase (Ca-ATPase), Vacuolar H⁺-ATPase (V-H-ATPase), F-type H⁺-transporting ATPase (F-H-ATPase), P-type H⁺-transporting ATPase (P-H-ATPase), Carbonic anhydrase (CA), chloride channel protein (Cl-channel), Na⁺-independent Cl/HCO₃ exchanger (Cl/HCO₃-exchanger), and innexin. b Expansion of F-ATPase in the E. sinensis genome. F-ATPase consists of two domains: F₀ domain (containing three subunits: a–c) that integrates in the membrane to promote passive H⁺ transport, and F₁ domain (containing five subunits: α, β, δ, γ, ε) which is easily released from the membrane, catalyzing hydrolysis of ATP⁴⁴. The expanded genes in each cluster share similar gene structures. The domains of each cluster were predicted by SMART. Genes from different species are represented by different colors: E. sinensis (red), P. hawaiensis (green), Daphnia pulex (orange), Drosophila melanogaster (blue). c The expression of osmoregulation related genes in megalopae (upper) and adults (lower) based on qPCR. Asterisks indicate the significant differences in gene expression between crabs in seawater (orange) and freshwater (blue). FHA F-H-ATPase, PHA P-H-ATPase, VHA V-H-ATPase, NKS Na/K-sympoter, CH Cl/HCO₃-exchanger. d Potential functions of F-ATPase in osmoregulation and development of E. sinensis. F-ATPase performs its function through regulating acid-base balance, energy expenditure, ion uptake, and estrogen-targeting in sex maturation.