JCS01315 Supplemental Figures

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Sequence analysis of distinct prominin-1 splice variants. (A) Comparison of nucleotide and deduced amino acid sequences of s1, s3, s5 and s6 variants (above) with that of the s4 variant (bottom) at the end of the first extracellular domain. A cryptic splice acceptor site (AG underlined) is present within the fifth exon. Arrowhead indicates an exon boundary in the prominin-1 gene. (B) C-terminal amino acid sequences of s3, s4, s5 and s6 variants deduced from the corresponding cDNA. Nucleotide and amino acid sequences identical to the s1 variant appear in blue, while the sequences of the s3, s4, s5 and s6 variants are shown in red. In the s6 variant, a premature termination codon (asterisk) is introduced due to the retention of an intron (black letters). Nucleotides matching the consensus for splice donor and acceptor sites (gt/ag) are indicated in small boxes, and two putative branch sites appear in bold. (C) Two polyadenylated prominin-1 cDNAs were amplified by 3’-RACE from mouse 11-day-whole embryo cDNA pool. Both contained part of the testis-derived EST clone 4932412G18 sequence encoding the end of s3, s4 and s5 variants (red letters) and 3’-UTR containing sequence identical to that of the s1 variant cDNA (nt 2731-3739) (green and blue letters). Nucleotide found in exons 24, 25a, 25b and 26 are delineated with boxes. The donor and acceptor splice sites (gt/ag) and the cryptic acceptor splice site (AG) are indicated in small boxes and the polyadenylation signal appears in bold. In the shorter cDNA, 431 nt long intron 25 (black lowercase letters) is spliced out.

• Supplemental Figure 2 -

  Fig. S2: Prominin-1 is not expressed in spermatogonia, spermatocytes and spermatids. Cryosections of adult murine testis (A,B) were immunolabeled with (A) or without (B) mAb 13A4 followed by peroxidase-coupled goat anti-rat and counterstained with Mayer’s hematoxyline. Spermatozoon precursors were identified according to the nuclei morphology. Asterisks, spermatozoon tails; arrows, spermatogonia; solid arrowheads, spermatocytes; open arrowheads, spermatids. Bars, 30 mm.

• Supplemental Figure 3 -

  Fig. S3: The C-terminal domain splicing cassette of mouse prominin-1 is conserved in rat  and human. The amino-acid sequences of the C-termini of s1, s3, s6 and s7 prominin-1 splice variants were aligned. a and b were deduced from the sequence of the respective rat and human genomic clones CH230-250 M20 and C0024K08 (GenBank Accession No. AC134757 and AC005598, respectively). The corresponding amino acid sequences appear in italic whereas sequences translated from cloned cDNA species identified with their GenBank Accession No. are in normal font.