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Figure 3 | BMC Molecular Biology

Figure 3

From: Characterization of the transcripts and protein isoforms for cytoplasmic polyadenylation element binding protein-3 (CPEB3) in the mouse retina

Figure 3

The expression of CPEB3 as demonstrated by RT-PCR. a) CPEB3 was present in the P60 retina. b) Multiple CPEB3 transcripts were present in the retina. Different primer sets for various CPEB3 transcripts (table1, table 2) were used for PCR. Each visible band was purified and sequenced, which confirmed the presence of six splicing patterns: "+69+24" (lane2, upper band), "-69+24" (lane2, lower band), "exon 11 extension" (lane3), "exon 11 + exon 12" (lane 4, upper band); a new pattern: "exon 11 deletion" (lane 4, lower band), and "partial exon skipping within exon 4". But it did not rule out the presence of "+69-24" and "-69-24", since the competition for the same set of primers by a dominant pattern ("-69+24") may mask the weakly expressed variants. c) Demonstration of "+69-24" and "-69-24" patterns and comparison of tissue distribution of all patterns. Primer sets specific for each individual splicing pattern were used for PCR on thirteen different tissues from adult mice. Each specific band was purified and sequenced for identity confirmation. The results demonstrated the presence of "+69-24" and "-69-24" patterns in the retina (lane 2) in addition to the 6 patterns identified in figure b). It also demonstrated the ubiquity of the majority of the patterns, with the exception of "+69+24", which was expressed in the CNS, the ovary, testis, kidney and heart, but not in the lung, liver, thymus and spleen. d) The locations of these primers were mapped to CPEB3. The corresponding relationships between the numeric primers and the splicing variants in figure a), b) and c) were listed in table 2. Darkened boxed represented exons that could be alternatively spliced.

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