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. 2018 Mar 23;9:1219. doi: 10.1038/s41467-018-03631-z

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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 — Histological score calculation based on the intensities of PLP staining and reproducibility of the staining. a Representative examples of the different PLP intensity levels in the cytoplasm and the nucleus of breast cancer FFPE tissue cells, compiled from different cases and different enriched libraries (×60, 10 µm scale bar). b Examples of the histological score calculations in two areas of the breast cancer tissue, stained with EL-NB. Note: assessment of the percentage of cancer cells with a particular staining intensity is usually performed on the entire tissue. Correspondingly, each stained case will have its unique cytoplasmic, nuclear and combined scores. Depending on the marker localization, nuclear, cytoplasmic or combined PLP scores can be informative for the differentiation of the benefiter from non-benefiter cases (×20, 20 µm scale bar). No adjustment was applied to images from microscopy. c Reproducibility of the PLP nuclear staining and scoring using EL-NB on eight different patients between technical replicates and different batches of the library preparation. TL-NB staining of the cytoplasm was not informative for the differentiation of B and NB cases. Patients 1–4 were selected as examples of relatively weak intensity staining, patients 5–8 are examples of relatively strong intensity staining (error bars are s.d. from three replicates)