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. 2017 Jul 14;7:5399. doi: 10.1038/s41598-017-05637-x

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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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Binding of cocaine increases the concentration of R60A (but not W63A) in the filopodia of HEK293 cells. (A) Cells transiently expressing wt YFP-HA-DAT, or its R60A or W63A mutants were incubated with 10 μM cocaine for 30 min at RT. Live-cell imaging was performed through the 515 nm (YFP, green) filter channel. Maximal z-projections of 5 consecutive x-y-confocal planes are shown. (B) Insets represent high magnification images of the regions marked by the white rectangle in (A). Arrows point to representative filopodia. (C) The filopodia/FM ratios of mean intensities of the YFP fluorescence were calculated as described in “Methods”. Results are shown as mean values (±SEM, n = 10). ***p < 0.001.