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. 2019 Sep 24;9:13753. doi: 10.1038/s41598-019-50276-z

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© The Author(s) 2019

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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Effect of LY-294002 on the expression of p-Akt and CLDNs. (A) The expression levels of p-Akt, Akt, p-PDK1, PDK1, p-ERL1/2, ERK1/2, p-c-Jun, and c-Jun in cell lysates were examined by western blotting. The levels of p-Akt, p-PDK1, p-ERK1/2, and p-c-Jun are shown as a percentage of the values in non-treated cells (Cont). (B) RERF-LC-AI cells were treated with or without 20 μM LY-294002 (LY) for 1 h. The expression levels of p-Akt are shown as a percentage of values in the absence of LY-294002. (C) The cells were treated with or without 20 μM LY-294002 for 24 h. The expression levels of CLDN1, CLDN11, and β-actin in cell lysates were examined by western blotting. The protein levels of CLDN1 and CLDN11 are shown as a percentage of values in the absence of LY-294002. The full-length blot images are shown in Supplementary Fig. S3. (D) The cells were treated with and without 20 μM LY-294002 for 6 h. The mRNA levels of CLDN1 and CLDN11 are shown as a percentage of values in the control (Cont). n = 3–4. **P < 0.01 and NS, P > 0.05 compared with the control cells.