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. 2018 Jun 1;37(39):5305–5324. doi: 10.1038/s41388-018-0294-0

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© The Author(s) 2018

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. 4 — Effects of stromal cell Dkk-3 on acinar morphogenesis and prostate cancer cell invasion. a Images of acini used for scoring. b Acinar morphogenesis (AM) assays (day 7) using shDKK3 (sh6) RWPE-1 cells cultured with assay medium (control), conditioned media (CM) from WPMY-1 cells or CM from shDKK3 (Wsh8) WPMY-1 cells; error bars show SD, n = 4, *p < 0.05, **p < 0.01. Similar results were obtained using shDKK3 NPSM and Wsh7 WPMY-1 cells and using shDKK3 (sh30) RWPE-1 cells (Supplementary Figures 6a and 6b). c AM assays (day 7) using shCTRL RWPE-1 (NS11) cells cultured with assay medium (control), CM from WPMY-1 cells or CM from shDKK3 (Wsh8) WPMY-1 cells; error bars show SD, n = 3, *p < 0.05; similar results were obtained using shCTRL (NS14) RWPE-1 cells with CM from shCTRL (NPSM) and shDKK3 (Wsh7) WPMY-1 cells (Supplementary Figure 6c). d Proliferation assays (48 h) using shDKK3 (sh30) RWPE-1 cells cultured in the upper chamber, separated by a 0.4-μm membrane from the lower chamber, which contained assay media or shCTRL (PSM2) or shDKK3 (Wsh8) WPMY-1 cells; n = 4, *p < 0.05. e Cell invasion assays using PC3 cells cultured with CM from parental WPMY-1 and shDKK3 (Wsh8) WPMY-1 cells, n = 3, *p < 0.01. Left, representative photos of invaded cells stained with crystal violet. Similar results were obtained using CM from shCTRL (NPSM) and shDKK3 (Wsh7) cells (Supplementary Figure 7). f Cell invasion assays using PC3 cells cultured with CM from shCTRL (NPSM) and shDKK3 (Wsh8) WPMY-1 cells with or without the MMP2 inhibitor ARP100; n = 3, ***p ≤ 0.001. Left: representative photos of invaded cells stained with crystal violet. Scale bars 100 μm