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. 2017 Oct 23;7:13816. doi: 10.1038/s41598-017-14124-2

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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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FTY720 promotes Rsp5-dependent endocytosis of multiple permeases. (A) Strains 23344C (ura3) and 27038a (rsp5/npi1 ura3) transformed with plasmid pJOD10 (YCpGAL-GAP1-GFP), pCJ563 (YCpGAL1-CAN1-GFP), pNAM001 (YCp-LYP1-GFP), or pFL38-gF-GFP (YCpGAL1-FUR4-GFP) were grown on galactose-proline (Gap1-GFP, Can1-GFP, and Lyp1-GFP cells) or galactose ammonium (Fur4-GFP cells) medium. Glucose was added for 90 min before addition of FTY720 or DMSO solvent alone (control). Cells were examined by wide field microscopy after a 2-h incubation. Cells were also labeled with CMAC dye to visualize the vacuole. (B) Strains 23344C (ura3) and 27038a (rsp5/npi1 ura3) transformed with an empty URA3 plasmid (pFL38) were grown on glucose ammonium liquid medium. FTY720 was added to half of the cultures. Relative growth rates were measured using strain 23344c in the FTY720-free medium as a reference (control). Values represent the means of three independent experiments and error bars correspond to standard deviations (SD) (unpaired t-test, *P < 0.05, ***P < 0.001).