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. 2012 Oct 4;153(12):6041–6053. doi: 10.1210/en.2012-1670

Fig. 1.

Fig. 1.

Expression of components of the Scribble polarity complex in cultured Sertoli and germ cells and the testis, and stage-specific localization of Scribble at the BTB during the epithelial cycle of spermatogenesis. A, Immunoblotting of Scribble in lysates of adult rat testis, Sertoli cells (SC), and germ cells (GC) with actin served as a loading control (50 μg protein/lane). B, RT-PCR results depicting the relative steady-state mRNA levels of Lgl2 and Dlg1 in lysates of testis, SC, and GC, with S-16 serving as a control. C, Histogram summarizes results of immunoblotting and RT-PCR shown in panels A and B after normalizing each data point against actin (for immunoblot) or S-16 (for RT-PCR). Protein (Scribble) or mRNA (Lgl2 or Dlg1) level in the testis was arbitrarily set as 1. Each bar is the mean ± sd of n = 3. D, Colocalization of Scribble (green) and basal ES protein β-catenin (red) to the Sertoli cell-cell interface by dual-labeled immunofluorescence analysis in cells cultured for 4 d. Nuclei were stained with DAPI (blue). Scale bar, 5 μm, which applies to other micrographs. E, Stage-specific expression and localization of Scribble at the BTB in the epithelium during the epithelial cycle. Scribble was expressed predominantly at the basal region near the basement membrane, consistent with its localization at the BTB, with the highest level in stages VII–VIII, at the time of BTB restructuring to facilitate the transit of preleptotene spermatocytes at the site. Negative control was shown in which the anti-Scribble antibody was substituted by normal goat IgG; scale bar, 40 μm, which applies to the micrograph on the right. Scale bar, 20 μm, in the staged I–II tubule, which applies to all other micrographs. F, Immunoblot analysis illustrating the specificity of the goat anti-Scribble antibody using Sertoli cell (SC) lysates. G, Colocalization of Scribble (green) with tight junction proteins occludin (red) or ZO-1 (red), predominantly at the BTB. Nuclei were stained with DAPI (blue). Scale bar, 30 μm, which applies to other micrographs.