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. 2009 Jan;150(1):251–259. doi: 10.1210/en.2008-0044

Fig. 4.

Fig. 4.

The defects of prostate development in the ACTB-Cre/ERα−/− males. A, Photographs demonstrating the branching morphogenesis from a representative pair of adult ACTB-Cre/ERα−/− and Wt littermates after microdissection of VP. B, The total number of ductal tips, branches, and ductal points in ACTB-Cre/ERα−/− and Wt littermates. The VP and DLP, but not AP, of ACTB-Cre/ERα−/− mice have significantly fewer ductal tips than those of WT littermates. *, P < 0.01 vs. Wt littermates (statistically significant by Student’s unpaired t test). Results are means ± sd (n = 5). C and D, The cell proliferation of VP from 1-wk-old ACTB-Cre/ERα−/− and Wt littermate is determined by Ki-67 immunostaining (C). Arrows show positive staining in the epithelial cells. Arrowheads show positive staining in the stromal cells. The Ki-67 proliferation index is scored as the percentage of Ki-67-positive cells (D). Note that there was significantly reduced stromal proliferation in the ACTB-Cre/ERα−/− VP. *, P < 0.05, compared with Wt VP cell proliferation, Student’s t test. The results are means ± sd of triplicate samples. E, Characterization of proliferating stromal cells in the ACTB-Cre/ERα−/− and Wt VP. Immunofluorescence analyses were carried out by using sections from neonatal VP lobes. Each section was double stained by Ki-67 and vimentin (fibroblast cell marker) or desmin (smooth muscle cell marker) followed by counterstaining with 4′,6′-diamino-2-phenylindole (DAPI) to visualize the nuclei. Arrowheads show positive Ki-67 staining in the fibroblasts (c, g, k, and o). Arrows show positive Ki-67 staining in the epithelial cells (c and g). Long arrow shows positive Ki-67 staining in the smooth muscle cells (k). Note the fibroblasts are actively proliferating (c and g), but the smooth muscle cells rarely proliferate (k and o) in the stromal compartment. F, The comparison of the expression of the stromal marker between ACTB-Cre/ERα−/− and Wt littermates by real-time PCR. Assays were performed on RNA from the VP of each individual mouse and then averaged for the gene expression. 18s acts as internal control. The results are means ± sd of six samples. Note that the expression of vimentin is decreased, but the expression of both smooth muscle α-actin and desmin is increased in the ACTB-Cre/ERα−/− VP. G, The comparison of the expression levels of the genes involved in prostatic branching morphogenesis between ACTB-Cre/ERα−/− and Wt littermates by real-time PCR. Assays were performed on RNA from the VP of each individual mouse and then averaged for the gene expression. 18s acts as internal control. The results are means ± sd of six samples. *, P < 0.05, compared with Wt gene expression, Student’s t test. Note that only BMP4 is significantly up-regulated in the ACTB-Cre/ERα−/− VP.