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. 2018 Nov 7;16:74. doi: 10.1186/s12964-018-0285-3

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© The Author(s). 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 4 — Hyperosmolarity increases Wnt/β-catenin signaling in mIMCD3 cells. a Expression levels of Wnt4, Wnt5a and β-catenin mRNA in mIMCD3 cells exposed to norm- or hyperosmotic media for 24-72 h measured by qPCR. The data obtained were normalized to the expression of the reference genes glyceraldehyde-3-phosphate dehydrogenase (Gapdh), β-actin (Actb), and β2-microglobulin (B2m). Means ± SEM of 3-4 experiments are shown. Statistical analysis compares the two osmotic conditions by unpaired t-test. b Expression of GSK-3β and phospho-GSK-3β (Ser9) in mIMCD3 cells was detected by immunoblotting. Optical density (O.D.) of phospho-GSK-3β (Ser9) and GSK-3β signals was analyzed by densitometry using ImageJ software, and relative O.D. was calculated from the ratio of phospho-GSK-3β (Ser9) / GSK-3β. Statistical analysis shows means ± SEM of 4 experiments and compares the two osmotic conditions by unpaired t-test.