Quantitative evaluation of the percentage of tissue area exhibiting CD34 or sma immunopositivity and of nuclear area exhibiting PR immunopositivity (LI, labeling index) in the ULP area.

<p>Each dot shows the value associated with one case. The results of the Kruskal-Wallis test and the post-doc pair comparison test are shown below.</p

Representative images of desmin-staining (marker for MM) showing the increased development of desmin+ MM in MS bladders compared to controls.

<p>MM is more developed in MS bladders, both vertically and horizontally. MM (indicated by red arrows) lies just below the ULP IC area (between the 2 dotted red lines). In BPS and CIS bladders there was a trend towards increased MM development, but no significant difference was found (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127020#pone.0127020.g003" target="_blank">Fig 3</a>). Scale bars indicate 600μm. U is urothelium, ULP IC equals upper lamina propria interstitial cells.</p

Table listing the details of the antibodies used in the present study.

<p>RTU equals ready to use.</p><p>Table listing the details of the antibodies used in the present study.</p

Representative images of staining for alpha-sma (marker for ULP IC) showing the increased thickness of the alpha-sma+ ULP IC area in MS and BPS bladders.

<p>The ULP IC area lies between the 2 dotted red lines. The alpha-sma+ cell-types under the ULP IC area are MM-fibres and perivascular smooth muscle cells (which were excluded from analysis); IC in the DLP are negative for alpha-sma. In BPS bladder heavy inflammatory infiltrate is laying in between the IC, while in MS bladder this infiltrate is less pronounced. U is urothelium, ULP IC equals upper lamina propria interstitial cell. Scale bars indicate 600μm.</p

Quantitative evaluation of the percentage of tissue area exhibiting CD34 or sma immunopositivity and of nuclear area exhibiting PR immunopositivity (LI, labeling index) in the ULP area.

<p>Each dot shows the value associated with one case. The results of the Kruskal-Wallis test and the post-doc pair comparison test are shown below.</p

Representative images of PR-staining, showing the decreased amount of PR+ ULP IC in MS bladders.

<p>PR shows a nuclear expression and is widely expressed in ULP IC. Asterisks indicate heavy inflammatory infiltrate in the ULP area in BPS. U is urothelium, ULP IC equals upper lamina propria interstitial cells (red dotted line separates urothelium form the ULP IC area). Scale bars indicate 300μm.</p

Representative images of CD34-staining (endothelial marker) showing the vascularisation of the bladder ULP area in the different studied groups.

<p>No significant difference was found in total vascularisation in any of the studied groups (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127020#pone.0127020.g005" target="_blank">Fig 5</a>). As shown in the images the vascular topography was changed in all bladder disease groups, with blood vessels being organised more haphazardly in the ULP, although this observation could not be quantified. Arrows indicate CD34+ blood vessels and capillaries. U is urothelium, ULP IC equals upper lamina propria interstitial cells (red dotted line separates urothelium form the ULP IC area). Scale bars indicate 300μm.</p

Top 5 upregulated genes in pairwise comparisons between tumor cells of NLPHL, THRLBCL-like NLPHL and THRLBCL.

<p>p-values <0.05. FDR (false discovery rate)<0.3. vs = versus.</p

Quantification of the microenvironment in typical NLPHL (patterns A and C), THRLBCL-like NLPHL (pattern E) as well as THRLBCL.

<p><b>a.</b> Numbers of CD4⁺ T cells/mm² in typical NLPHL (pattern A: n = 14 and pattern C: n = 13), THRLBCL-like NLPHL (n = 14) and THRLBCL (n = 25). (*p<0.05, **p<0.01, unpaired t-test). <b>b.</b> Numbers of CD8⁺ T cells/mm² in typical NLPHL (pattern A: n = 14, pattern C: n = 15) and THRLBCL-like NLPHL (n = 12) as well as THRLBCL (n = 22). <b>c.</b> Numbers of CD163⁺ macrophages/mm² in NLPHL (pattern A and C as well as THRLBCL-like NLPHL: n = 14, each) and THRLBCL (n = 25), (***p<0.001, Mann-Whitney-test).</p

Immunophenotype of rosetting T cells in different variants of NLPHL and THRLBCL^*.

*<p>Cases were scored positive if positive rosetting T cells were observed around at least 5% of the tumor cells.</p