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. 2017 Jul 26;8:142. doi: 10.1038/s41467-017-00084-8

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© The Author(s) 2017

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. 7 — AIM1 and actin localization in normal prostate, prostate adenocarcinoma and prostate embryonic development. a Co-immunolabeling of AIM1 (red) and β-actin (green) reveals high degree of co-localization in normal prostate epithelium. In prostate cancer, this co-localization is largely abolished and AIM1 shows a diffuse cytoplasmic staining pattern. Nuclei are counterstained with DAPI. b Boxplot shows distribution of co-localization coefficients of AIM1 and β-actin immunostaining in prostate cancer and adjacent benign prostate epithelium (n = 8 cases). Scale bars indicate 50 μm. c Proximity ligation assays (PLA) confirm close spatial proximity of AIM1 and β-actin in benign prostate epithelial cells and greatly reduced interaction in invasive carcinoma. Red dots indicate discrete interaction signals by PLA; green shows β-actin immunostaining. Scale bars indicate 100 μm. d Boxplot shows differences in area-normalized PLA signals in benign glands and invasive carcinoma (n = 8 cases). e Immunolabeling for Aim1 (shown in red) and β-actin (shown in green) in urogenital mesenchyme (UGM) and urogenital sinus epithelium (UGE) from mouse embryos at day 17.5 post conception. Note the UGE-specific expression of Aim1, with membranous staining and actin co-localization in bulk UGE (arrowheads), and more diffuse cytoplasmic staining in UGE-derived prostatic buds (labeled B) invading into the surrounding UGM (arrows). Scale bars indicate 100 μm. f Boxplot shows distribution of co-localization coefficients of Aim1 and β-actin immunostaining in UGE close to the urogenital sinus (Bulk UGE) and invading prostatic buds (Bud) (n = 4). g IHC confirms membranous staining pattern of Aim1 in bulk UGE and decreased, diffuse Aim1 immunoreactivity in invading buds. h PLA demonstrates spatial proximity of Aim1 and β-actin UGE close to the urogenital sinus and decreased PLA signals in invading prostate buds. Scale bars indicate 100 μm. i Boxplot shows differences in area-normalized PLA signals in bulk UGE and invading buds (n = 3). All P values are derived using t-test statistics