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. 2019 Jan 29;9:857. doi: 10.1038/s41598-018-37408-7

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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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Spatial scores of iron deposits from Prussian blue histology in human breast cancer. Prussian Blue iron histochemistry shows the presence of iron deposits in (a) stromal margins of carcinoma in situ (CIS) and (b) invasive carcinoma (INV, blue arrows). No iron deposits were associated with (c) invasive carcinoma exhibiting poorly defined stromal margins. Scale bar 200 µm. Expansions of boxes in (a–c) shown below. Scale bar 40 µm. Concentric rake region of interest grid overlay used to profile HLMs in (d) CIS and (e) INV fields. Scale bar 100 µm. (f) Iron⁺ macrophage (HLM) infiltration profiles from Prussian blue histology in CIS and INV fields. (mean + s.e.m. n = 5 fields/cancer subtype, **p < 0.01, ****p < 0.0001, 2-way ANOVA with Sidak’s multiple comparison test).