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. 2019 Mar 20;9:4954. doi: 10.1038/s41598-019-40848-4

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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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Selective blockade of EP4 significantly reduces tumor sphere formation of HT29 OXR cells. The effect of EP4 inhibition on tumorigenicity was tested in HT29 OXR cells. (A) Untreated PAR and OXR cells were plated in an ultra-low attachment 96-well plate (100 cells per well) for 1 week and the numbers of viable tumor spheres per field were counted. (B) HT29 OXR cells were treated with PTGES siRNA or non-targeting (NT) siRNA for 48 hours, followed by a tumor sphere formation assay (Materials and Methods). Columns, means of samples of two independent experiments; bars, SEM. **P < 0.01, ***P < 0.001 (Student’s t-test). (C) OXR cells were seeded in an ultra-low attachment plate and treated with the indicated combination of oxaliplatin and the EP-4 inhibitor, L-161,982 for 1 week. At day 7, the number of viable tumor spheres per field were counted. Columns, means of samples of two independent experiments; bars, SEM. ***P < 0.001 (One-way ANOVA).