Fig. 2. TIMP-2 mRNA contains DDC8 sequence.
A) EST database analysis indicating potential alternative splicing between DDC8 and its host gene TIMP-2. B) First strand cDNA was primed either with oligo (dT) or with an oligonucleotide specific to TIMP-2 exon 2 (T2). The cDNAs were then amplified with primers within DDC8 exon 3. The oligo (dT) cDNA demonstrates that DDC8 is expressed in adult TIMP-2-/- (KO) and wild-type (WT) brain. Amplification of the TIMP-2 primed cDNA reveals the presence of TIMP-2 transcripts containing DDC8 sequence in adult TIMP-2-/- and wild-type mice, and rat brain. TIMP-2 primed cDNA in the absence of reverse transcriptase (no RT) fails to amplify a DDC8 exon 3 product, suggesting the products did not arise from contaminating genomic DNA. C) PCR with oligo (dT) primed adult cDNA amplifies the three DDC8 exons in testis, but only exon 3 in brain indicating that DDC8 mRNA expressed in brain differs from that expressed in testis.