Mean absolute expression of human USP26 protein in an array of human normal and neoplastic tissues.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098638#pone-0098638-t002" target="_blank">Table 2</a> demonstrates USP26 protein expression in non-testicular tissue including androgen-dependent cancers. Expression ratio is defined as higher expression divided by lower expression. Cancer versus normal column is directional change showing increased or decreased in cancer relative to normal tissue from each organ (+ =  increase as compared to normal tissue; − =  decrease in expression compared with normal tissue). Expression ratio then used to classify expression pattern (Green =  overexpression; White =  ratio +2 to −2 (equivocal expression), Red =  lower expression compared to normal tissue).</p

USP26 and AR protein expression in breast and thyroid tissue by immunofluorescence in formalin-fixed, paraffin-embedded human tissue.

<p><b>4A</b>: USP26 staining in benign breast tissue (paraffin-embedded fixed), mostly cytoplasmic with some nuclear staining in secretory luminal cells (right arrow) and myoepithelial cells (left arrow). <b>4B</b>: USP26 staining in benign thyroid follicular cells (arrows), mostly cytoplasmic localization.</p

USP26 pF1K and pFN2A GST Flexi Vectors.

<p>Adapted from pF1KT7 Flexi Vector map with sequence reference points (Promega: Madison, WI)</p

USP26 and AR protein expression in normal testis by immunofluorescence.

<p>All images of stained sections were captured at 400x total magnification; scale bar = 100 µm. <b>3A</b>: Colocalization of USP26 and AR in nucleus and cytoplasm of Leydig cell (A), spermatogonia (B), primary spermatocyte (C), and Sertoli cell (D) in normal human testis (frozen tissue). <b>3B</b>: Partial colocalization of USP26 and AR in nucleus and cytoplasm of Sertoli cell (left arrow) and spermatogonia (right arrow) of normal human testis (frozen tissue). <b>3C</b>: Colocalization of USP26 and AR in nucleus and cytoplasm of Sertoli cells (left arrow) and spermatogonia (right arrows) of normal human testis (frozen tissue). <b>3D</b>: USP26 expression in spermatids (double arrows), and USP26/AR partial colocalization in early cells of spermatogenesis (single arrows) of normal human testis (frozen tissue). <b>3E</b>: Negative control demonstrating specificity of antibody without staining of USP26 or AR, or auto-fluorescence of normal human testis (frozen).</p

USP26 pF1K and pFN2A GST Flexi Vectors.

<p>Adapted from pF1KT7 Flexi Vector map with sequence reference points (Promega: Madison, WI)</p

Gel Electrophoresis demonstrating USP26 mRNA expression in thyroid and breast cancer tissue following qRT-PCR.

<p>USP26 mRNA expression in breast and thyroid cancer confirmed by gel electrophoresis with amplicon for USP26 noted in breast cancer, thyroid cancer, and normal testis.</p

Testosterone production in human testis biopsy after 3 hours.

<p>Experiments in humans were performed using 12 testis samples. All results were pooled in statistical analysis using one-way Anova.</p

Dose response analysis of testosterone production in rat Leydig cells with E2, G-1 and ICI incubation.

<p>Final concentration of 10, 50, 100 and 500-statistically significant difference between 10, 50 and 100 nM.</p

MTS viability assay performed with different concentrations of G-1, after 3 and 12 hours.

<p>Data was analyzed using two-way ANOVA with time and concentrations as factors.</p

Localization studies of GPER-1 in rat testis.

<p>A. Immunohistochemistry imaging of adult rat testis. GPER-1 antibody is labeled in red. B. Immunofluorescence localization analysis of adult rat testis, with GPER-1 labeled in red, and nuclei conterstained with DAPI (blue). C. High resolution Z-stack imaging with 3D deconvolution obtained from each fluorescent channel using motorized microscope.</p