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. 2021 Mar 17;12:1714. doi: 10.1038/s41467-021-21976-w

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© The Author(s) 2021

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 — a, b Bone metastatic growth of PCa cells (DU 145, PC-3, and C4-2B) expressing different forms of KLF5, including tumor formation rate (a) and radiographs (b) at 5 weeks (DU 145 and PC-3) or 12 weeks (C4-2B) after tibial inoculation of cells. PCa cells with KLF5 knockout (KLF5 null) were infected with lentiviruses to express empty vector (EV), wild-type KLF5 (KLF5), KLF5^K369R (KR), and KLF5^K369Q (KQ). White arrows point to bone lesion areas. c H&E staining of tibial tumor samples from PCa cells with indicated forms of KLF5. B trabecular bone regions, BM bone marrow regions, T tumor regions. d Statistical analysis of the images in c by calculating the ratio of tumor area to total sample area of DU 145 (left), PC-3 (middle), and C4-2B (right) cells with different forms of KLF5 in bone. For DU 145 cells, n = 3, 7, 4, and 8 tumors for EV, KLF5, KR, and KQ respectively. For PC-3 cells, n = 6 tumors for each group. For C4-2B cells, n = 8 tumors for each group. e, f IHC staining (e) and intensity quantification (f) of epithelial marker E-cadherin and mesenchymal marker vimentin in tibial tumors of DU 145 and PC-3 cells with different forms of KLF5. For DU 145 cells, n = 3, 3, 4, and 5 tumors for EV, KLF5, KR, and KQ respectively. For PC-3 cells, n = 3, 4, 5, and 5 tumors for EV, KLF5, KR, and KQ respectively. g, h IHC staining (g) and signal intensity quantification (h) of proliferation marker Ki67 in tibial tumors of DU 145 (Left) and PC-3 (right) cells. n = 3 tumors for each group of PC-3 cells. For DU 145 cells, n = 2, 6, 3, and 6 tumors for EV, KLF5, KR, and KQ respectively. One representative field per tumor was used for statistical analysis in e–h. Scale bar, 50 μm. In panels d, f, and h, data are shown in mean ± S.E.M. NS not significant; *p < 0.05; **p < 0.01 (two-tailed Student’s t test). The Fiji software was used to quantify staining signal intensities in panels f and h. Source data are provided as a Source Data file.