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. 2017 May 3;7:1390. doi: 10.1038/s41598-017-01500-1

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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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Heatmap of gene expression profiles obtained by treatment of WY-14643. The dendrogram was made without log-transformation of the axis, according to a hierarchical clustering result (method: ward.D2, distance: Pearson). (A) Gene expression profile of M4vs8_cluster1 (probe: n = 268). The x-axis represents experimental conditions (from left to right, (1–8) WY-14643 3 hr, 6 hr, 9 hr, 8 day, 15 day, 29 day, 24 hr and 4 day; (9–11) amlodipine 6 hr, 24 hr and 3 day. The y-axis represents probes in M4vs8_cluster1. (B) Gene expression profile of M4vs8_cluster2 (probe: n = 188). The x-axis represents experimental conditions (from left to right): (1–3) amlodipine 24 hr, 6 hr and 3 day; (4–11) WY-14643 4 day, 3 hr, 6 hr, 15 day, 29 day, 8 day, 24 hr and 9 hr. The y-axis represents probes in M4vs8_cluster2. (C) Gene expression profile of M4vs8_cluster3 (probe: n = 162). The x-axis represents experimental conditions (from left to right, 1–7) WY-14643 29 day, 15 day, 8 day, 3 hr, 6 hr, 24 hr and 9 hr; (8–10) amlodipine 24 hr, 3 day and 6 hr; 11) WY-14643 4 day). The y-axis represents probes in M4vs8_cluster3. (D) Analytical workflow for obtaining three M4vs8_clusters in (Fig. 2A–C). A gene set was selected by filtering DEGs between WY-14643/M dose/4 day and WY-14643/M dose/8 day by Welch’s t-test (cutoff: p-value = 0.01). As columns, Open TG-GATEs samples of WY-14643 (single dose (30 mg/kg bw): 3 hr, 6 hr, 9 hr, 24 hr, repeat dose (30 mg/kg bw): 4 day, 8 day, 15 day, 29 day) and external samples of amlodipine (uploaded to Toxygates by “user data upload function”, L dose (0.2 mg/kg), 6 hr or 24 hr, and M dose (19 mg/kg), 3 day) were used. The clusters were obtained by hierarchical clustering (dendrogram cutoff: 6). The high-resolution image file is available at Scientific Reports online.