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. 2017 Sep 7;7:10765. doi: 10.1038/s41598-017-11370-2

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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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Labelling of 5-HT_2B receptors by compound 12i in cells. (A) Living parental untransfected CHO-K1 cells (CHO-K1) and CHO-K1 cells stably expressing 5-HT_2B receptors (CHO-K1-5-HT_2B) were incubated in the absence or presence of the indicated concentrations of compound 12i and, after compound removal, fluorescence images (excitation wavelength 520–550 nm, emission wavelength 560–630 nm, standard filter set for 5-TAMRA) were acquired using an automated high content imaging instrument at 20x magnification. (B) Sample images from CHO-K1 and CHO-K1-5-HT_2B cells labelled with 1 µg/mL Hoechst 33342 (for nuclear staining) and compound 12i at a concentration of 3 μM, sampled from those quantified in (C,D). Minimum and maximum intensity and gamma correction of the images are shown in the colour scale in the panel. (C,D) Quantification of the fluorescence emission of compound 12i in images from CHO-K1 and CHO-K1-5-HT_2B cells labelled with 1 µg/mL Hoechst 33342 (for nuclear staining) and compound 12i at a concentration of 3 μM, both by image analysis (C) and by direct fluorescence measurement using a plate reader (D). The graphs show mean ± SEM of 4 wells, 5 fields/well (image analysis) and mean ± SEM of the same wells (plate reader). ***p < 0.001, two-way ANOVA and Bonferroni post tests.