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. 2021 Feb 1;12:723. doi: 10.1038/s41467-021-20962-6

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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. 4 — a Parental (F0 PAIII) and MSC selected (F2 PAIII) growth over time in the presence (1:1 ratio) or absence of MSCs (n ≥ 6/group). Representative images of bioluminescence in each group are shown at day 11 time point. Graphs illustrate collected RLUs over time for each group. Non-linear regression was used for statistical analysis with mean ± SEM, p = 0.0052. b Representative images (n≥3 biologically independent samples) of smooth muscle actin staining (α-SMA; red) in tissues derived from the F0 and F2 groups in the presence or absence of MSCs. Pan-cytokeratin (pCK; green) was used to localize prostate cancer cells. Dashed box in merge represents area of magnification. c, d Ex vivo analyses from study endpoint of proliferative and apoptotic indices using phospohistone H3 (pHH3; red arrows; b) and cleaved caspase 3 (CC3; red, arrows, b), respectively. Pan-cytokeratin (green) was used to identify prostate cancer cells (n ≥ 3 fields of view from at least three independent experiments). e μCT scan analysis of cancer-induced bone destruction. Representative μCT images of the trabecular bone are shown for the F0 and F2 PAIII group. The trabecular bone volume was calculated as a ratio to total volume analyzed (BV/TV; n ≥ 3 bones from each group). f Trabecular bone volume (BV) was measured via histomorphometry on non-sequential H&E multiple sections derived from each group and calculated as a percentage of total volume (n ≥ 4 bones derived from each group). Representative gross H&E images are illustrated from the F0 and F2 groups. g The number of osteoclasts (TRAcP positive; red, multi-nucleated; arrows) per μm of bone was calculated in non-sequential sections derived from each group (n ≥ 3 fields of view from at least three independent experiments). For all graphs shown, error bars represent mean ± SEM, statistical analyses were performed by one-way ANOVA with multiple comparisons. Asterisks denotes statistical significance (*p ≤ 0.05, ****p ≤ 0.0001) while NS denotes not significant.