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. 2018 Feb 21;8:3398. doi: 10.1038/s41598-018-21768-1

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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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DpdtC up-regulated NDRG1 expression and down-regulated membrane expression of HER2 on SK-OV-3 cells. (A) SK-OV-3 cells were incubated for 24 h at 37 °C with control media (−) or media containing the DpdtC (2 μM), and NDRG1 level was examined by western blot. And quantification of western blot signal intensity analysis is expressed relative to the β-actin loading control by using Image J software. ***p < 0.001. (B) Representative micrographs of fluorescent immunostaining showed that DpdtC decreased HER2 expression at the cell membrane. Cells were incubated for 36 h at 37 °C with control media (−) or media containing the DpdtC (2 μM), and HER2 localization was examined via immunofluorescence. Original magnification, ×600. Scale bars, 25 μm. (C) Percentage of HER2-positive stained cells reduced markedly in SK-OV-3 cells when incubated with DpdtC. Data are expressed as mean ± SD of the integrated fluorescence signals from 3 fields for each specimen. (D) Flow cytometry assay quantifying the membrane expression of HER2 on SK-OV-3 cells, which was expressed as MFI (Median Fluorescence Intensity) upon treatment control media (−) or media containing the DpdtC (2 μM) for 36 h at 37 °C. **p < 0.01; ***p < 0.001.