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. 2018 Nov 8;8:16551. doi: 10.1038/s41598-018-34877-8

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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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Cellular uptake of rapamycin analogues in VSMCs. (a) Chemical structures of three rapalogues- sirolimus (Siro), everolimus (Ever), and biolimus (Bio). Structures are drawn using ChemDraw software (PerkinElmer). Different moieties attached to C40 of the common macrolide core are shown. (b) Elution profile of rapalogues from high-performance liquid chromatography. The indicated rapalogue in methanol (10 μM) was separated on C₁₈ column and then eluted by 50–95% acetonitrile gradient in 0.1% trifluoroacetic acid solution. The chromatogram shows the different retention times and absorbance intensities of the drugs measured at 278 nm. (c) Quantification of the concentration of rapalogue absorbed in HASMCs. The methanol extracts from cultured cells treated with rapalogues (2 and 10 μM in PBS) for 2 hr were separated on C₁₈ column as in (B). Concentration was calculated from a standard curve of each compound. Bar in the graph are means ± SD of intracellular concentrations of rapalogues (n = 3, ^*P < 0.05 with Student’s t-test). N.S., not significant.