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. 2018 Oct 24;8:15726. doi: 10.1038/s41598-018-33879-w

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© The Author(s) 2018

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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Fisetin mediated decrease in RSK/YB-1 does not require suppression of ERK signaling (A) Whole cell lysates of A375 cells treated with fisetin (40–60 µM:24 h) were analyzed for ERK1/2 and RSK protein expressions. Equal loading was confirmed by reprobing for vinculin. (B) WM35 melanoma cells, transfected with pcDNA-HA-YB-1, treated with fisetin (60 µM:24 h) were analyzed for ERK1/2 expression. Equal loading was confirmed by reprobing for vinculin. (C) Whole cell lysates of NCI/ADR-Res ovarian cancer cells treated with fisetin (40–80 µM:24 h) were analyzed for phosphorylated RSK and ERK1/2 protein expressions. Equal loading was confirmed by reprobing for vinculin. (D) Representative images (20x) showing immunofluorescence for p-RSK (red) in fisetin-treated (60 µM:24 h) NCI/ADR-Res cells. Scale bar, 20 µm. (E,F) Whole cell lysates of A375 melanoma and NCI/ADR-Res ovarian cancer cells, treated with fisetin (60 µM:24 h) and/or ERK1/2 inhibitor FR180204 (10 µM) were analyzed for phosphorylated RSK and YB-1 protein expressions. Equal loading was confirmed by reprobing for vinculin. Data shown are representative of three independent experiments with similar results.