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. 2017 Jun 23;7:4135. doi: 10.1038/s41598-017-04509-8

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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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Glucocorticoid receptor (GR) mediates the effects of prednisolone. hCECs were cultured with or without 10 μg/ml prednisolone for 3 days and subjected to assays. (A) Real-time RT PCR assay for GR expression. (B) Representative image and densitometry analysis of western blot for GR relative to β-actin. (C,D) MTT assay and BrdU uptake assay to measure cell metabolic activity and proliferation, respectively. (E–G) Flow cytometric analysis for ANX⁺PI⁺ cells to measure cell apoptosis. The GR antagonist RU-38486 was added to some cultures (E). The percentage of ANX⁺PI⁺ cells was measured in hCECs with GR siRNA or scrambled (SCR) siRNA transfection (F,G). Data are presented as mean ±/+ SD from five independent sets of experiments including at least five samples per a set. **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: not significant.