Fig. 3. Changes in C5-530a2 mRNA expression during early postnatal development of the rat ovary, as assessed by an RNase-protection assay. (A) Above: the hypothetical gene (BC031519) predicted to encode a 317 amino acid protein; beneath: C5-530a2 (NM_184050) that was demonstrated experimentally to have a longer open reading frame. The predicted coding regions are depicted in gray and the untranslated regions in black. The approximate locations of the two probes used for RNase-protection assay are shown as white rectangles. The two putative ATG sites are indicated. (B) Left panel shows a representative autoradiogram depicting the increase in C5-530a2 mRNA abundance (detected by RNase-protection assay using probe A) between F21 and PN 48 hours, and the decrease towards adult values seen thereafter. Probe A consists of 289 nt transcribed from a C5-530a2 cDNA template plus 88 nt derived from transcribed vector sequences. The cyclophilin probe (Cyclo) is 211 nt in length, of which 158 nt correspond to transcribed vector sequences. The mRNA species protected by probe A, and the cyclophilin cRNA probe, are arrowed. MM, molecular weight markers (³²P-labeled RNA ladder); UP, undigested probe; DP, digested probe; A, adult ovaries. Right panel is a densitometric analysis of the changes in C5-530a2 mRNA levels detected by RNase-protection assay. RNA abundance is expressed as arbitrary units (AU) calculated using the individual C5-320a2/cyclophilin mRNA ratios from each sample. ^**, P<0.02, 48 hour group versus all other groups. Bars are mean values for each group and vertical lines represent s.e.m. (C) The same RNase-protection assay analysis as in B, but using probe B and samples pooled from selected developmental ages. Probe B consists of 492 nt, of which 406 correspond to transcribed vector sequences.