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. 2019 Jun 25;121(2):139–149. doi: 10.1038/s41416-019-0498-2

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© The Author(s) 2019

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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Fig. 2 — Crenolanib induces nuclear aberrations and apoptosis. a DAPI staining of A2780, ECRF24, HUVEC and HDFa after treatment with crenolanib for 72 h. In cells treated with crenolanib 5 µM white arrowheads indicate either micronuclei or undivided nuclear doublets. b, c Quantification of the percentage of micronuclei (b) and undivided nuclear doublets (c) based on images of DAPI staining. Values shown represent the percentage aberrations of the total amount of cell nuclei per image field. d Propidium-iodide staining of HUVEC treated with crenolanib for 72 h using flow cytometry. Cellular DNA content in permeabilised cells is proportional to fluorescence intensity (FL2-H; y-axis) and allows for the distinction of cells containing diploid DNA (G0/G1), tetraploid DNA (G2/M) and subdiploid DNA (apoptotic fraction). e Quantification of cellular DNA distribution over cell cycle phases as indicated in HUVEC, ECRF24 and A2780 cells after crenolanib treatment. All values shown are presented as percentage of the CTRL and represent the mean of at least two experiments. Error bars indicate SEM. Significance (*P < 0.05, **P < 0.01) is indicated as compared to control