Northern blot analysis and expression kinetics of the <i>R53</i> transcript during testicular development.

<p><b>(</b>A) A total of 10 μg of total RNA was extracted from each of the adult (4-month-old; lane 1) and immature testes (P14; lane 2) of the wild-type mice and the <i>Mvh</i> homozygote adult testis (lane 3) and run using Urea-PAGE. Then, the electro-blotted membranes were hybridized with [³²P]-labeled ASpB1D probe or AS2 probe. (B) Single-stranded cDNA was prepared from the testes on the indicated days after birth. The expression levels of the <i>R53</i> and selected spermatogenic stage-specific genes (including <i>Dmc1; DNA meiotic recombinase 1</i> and <i>Prm1; protamine1</i>) were quantitatively analyzed with <i>R53</i>-S1 and gene-specific primer pairs (listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0179585#pone.0179585.s010" target="_blank">S1 Table</a>). The values indicated are the relative values of the expression levels of each gene in the adult (4-month-old) testis (set as 1.0) after standardization to the <i>β-actin</i> expression level in the testes. The error bars indicate the SEM (n = 6).