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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 elevated the liposome permeability through a PS-related mechanism. The ITC data suggested the direct binding of the positively charged L-K6 to the negatively charged PS (a). Consistent with the data from the cell-based system, co-incubation of L-K6 with the liposomes caused rapid leakage of calcein from the liposomes in a dose-dependent manner. In addition, this leakage activity was PS-associated, as shown by an elevated calcein intensity with increased PS content of liposome (from 9:1:1 to 7:3:1 or 5:5:1) (b). Interestingly, calcein and carboxyfluorescein (CF) were released from the liposome, whereas FD10 and FD20, two fluorescent dyes with a molecular weight over 10kD, were resistant to be released (c). Moreover, the higher ratio of PS content in liposome yielded a higher percentage of α-helical structure, as assessed by circular dichroism spectroscopy (d). All experiments had been performed in triplicate. All data were expressed as mean ± SD.