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Fig. 2. Cell-type-specific expression of the gene encoding NO145 and alterations of the mRNA poly(A) tail during maturation. (A) RT-PCR analyses of NO145 and NO38 expression from various tissues of X. laevis (lanes 1-4: ovary, testis, muscle, and heart, respectively). (B) Scheme of the RACE-PAT assay adapted from Sallés et al. (Sallés et al., 1999). Total cellular RNA was reverse-transcribed using an oligo d(T) primer fused to a G/C-rich anchor sequence. Subsequent PCR amplification with a NO145-specific primer and the oligo d(T) anchor yielded a mixture of PCR-products representing the length of the poly(A) tail. (C) Top: ethidium bromide-stained PCR products obtained by the RACE-PAT assay on total RNA of X. laevis staged oocytes (I-VI) and eggs (E). Bottom: immunoblot of SDS-PAGE-separated total proteins from one oocyte of stages I–VI and from an unfertilized egg, per lane, and probed with NO145 and Xp-54 antibodies. (D) Alignment of oocyte and egg NO145 gene transcripts amplified by the RACE-PAT assay. Vertical lines represent identical residues between the transcripts. The stop codon is underlined and the nuclear polyadenylation signal is boxed. Poly(A) stretches are shown in bold letters.

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