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. 2017 Aug 15;7:8293. doi: 10.1038/s41598-017-08963-2

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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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L-K6 at higher concentrations preferentially elevated cancer cell membrane permeability. To evaluate the possible impacts of L-K6 on cell membrane permeability, a calcein/EthD-1 leakage assay was performed. L-K6 administration (especially 30~50 μM) induced a significant change in membrane permeability of MCF-7 cells, as indicated by the decreased fluorescence intensities of calcein (a) and increased EthD-1 (b). Additionally, L-K6-induced elevation of membrane permeability was cancer-specific, as evidenced by stable fluorescence intensities of calcein and EthD-1 in HaCaT cells (a,b). Consistent with calcein/EthD-1 leakage assay, the membrane depolarization assay further supported the elevated membrane permeability (mean ± SD) as indicated by the increased DiBAC₄(3) fluorescence intensity after peptides exposure (c). Morphological changes of human MCF-7 breast cancer cells induced by L-K6 as assessed by scanning electron microscopy (d) and transmission electron microscopy (e). *p < 0.05, **p < 0.01 vs. untreated control group. All experiments had been performed in triplicate.