XB-IMG-137173
Xenbase Image ID: 137173
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Figure 5. Decreased expression of Nkx3.1 in the Bmpr1a-CKO mutants. A and B, Nkx3.1 expression decreased in mutant prostate at P1 (A) and in the mutant AP at P28 (B). The relative mRNA equivalents for each sample were normalized by the RNA levels for ribosomal protein L8. Bars represent the mean ± SE of triplicate assays of RNA from pooled tissues (A) and 6 tissue samples (B). Statistical significance was indicated by asterisks (*, P < .05). C, Nkx3.1 was detected in the luminal epithelia of the control AP at P28. D, Significantly reduced levels of Nkx3.1 protein were detected in the mutant AP. E, The ratios of Nkx3.1-positive cells were shown in the graph. F, Colocalization of Nkx3.1 and Bmpr1a in the AP luminal epithelia of adult mice. G, Colocalization of pSmad1/5/8 and Nkx3.1 in the AP luminal epithelia of adult mice. F and G, Cryosections were used. Scale bars, 20 μm. H, Genomic sequences of the mouse Nkx3.1 aligned with its orthologous loci in human and opossum. The sequence alignment was performed using MultiPipMaker. A noncoding region conserved from human to opossum was indicated with black boxes in the C1 and C2 regions. The black arrow indicated exons of mouse Nkx3.1. Coding and untranslated sequences were shaded with red and yellow, respectively. A 5-kb (5399 base) region in the 3′-genomic region of Nkx3.1 contained a candidate enhancer region for the mouse prostate. The scale at the bottom of the alignment indicated relative positions in the mouse Nkx3.1 locus. I, The candidate 5-kb prostatic regulatory enhancer activated expression of a luciferase reporter in response to Smad1/4 expression (by 6 independent assays). It also responded to the addition of Bmp7 (by 3 independent assays) (means ± SE) (*, P < .05). a, Control. b, Transfected with Smad1/4 gene. c, Control. d, Transfected with Smad1/4 gene + addition of Bmp7. J, ChIP/PCR assay on bladder neck of ICR mice including prostate region at P2 showed pSmad1/5/8 binding to regions of C1 and C2 in the 3′-region of mouse Nkx3.1. Both regions were enriched in chromatin immunoprecipitated with antiacetylated histone H3 as a positive control. Image published in: Omori A et al. (2014) Copyright © 2014 by the Endocrine Society. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |