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. 2017 Sep 11;7:11210. doi: 10.1038/s41598-017-10348-4

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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/.

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Confocal images of SNARF-1 labeling of the calcifying medium of biominerals and pH measurements in different intra- and extracellular compartments. Transmitted light images of dumbbell-shaped crystals deposited on the coverslip (A) and a growing sclerite in the mesoglea (A’). (B-B’) Fluorescence images of SNARF-1 labeling showing the presence of an extracellular calcifying medium around biominerals. CEM: Cristal Extracellular Medium; SEM: Sclerite Extracellular Medium. (C) In vivo pH measurements obtained in cells and calcifying extracellular medium of C. rubrum microcolonies. Data are compared to the seawater pH (SW) surrounding microcolonies. Data are mean per microcolony ± standard deviation. SW: Seawater (n = 7); GCF: Fluid of gastrodermal canals (n = 4); CEM: Crystals extracellular medium (n = 5); AE: Aboral epithelium (n = 4); SEM: Sclerites extracellular medium (n = 4); Sclb: Scleroblasts (n = 7). Letters indicate mean pH values significantly different (one-way ANOVA with Leven’s post-hoc).