FIGURE

Fig. S6

ID
ZDB-FIG-131105-45
Publication
Andersson et al., 2012 - Adenosine Signaling Promotes Regeneration of Pancreatic beta Cells In Vivo
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Fig. S6

Test of the a2aa splice-blocking morpholino by RT-PCR and sequencing of truncated transcript, related to Figure 5.

(A) RT-PCR showing reduced amount of spliced a2aa mRNA in larvae injected with 5 or 10 ng a2aa MO at 4, 5, and 6 dpf. 10 larvae from each condition and stage were used for cDNA synthesis. The a2aa morpholino blocks the splice donor site of the first coding exon (dashed underlined in red) leading to a decreased amount of PCR product as the primers anneal to the first and second exons (underlined sequence). A truncated transcript appeared in the presence of the MO. Equal amounts of cDNA template are indicated by amplification of elongation factor 1-α, using the primers (5′-tcaccctgggagtgaaacagc-3′) and (5′-acttgcaggcgatgtgagcag-3′). (B) Sequence of the normal a2aa cDNA and splice junction (black /). The truncated transcript formed in the presence of the splice blocking MO used an alternative splice junction (red /), leading to the deletion of the sequence labeled in red. This deletion causes a frameshift that creates a premature stop codon (double underlined), thereby leading to a truncated protein of 149 amino acids that only includes 55 out of the 442 amino acids of the full-length A2aa protein.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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Reprinted from Cell Metabolism, 15(6), Andersson, O., Adams, B.A., Yoo, D., Ellis, G.C., Gut, P., Anderson, R.M., German, M.S., and Stainier, D.Y., Adenosine Signaling Promotes Regeneration of Pancreatic beta Cells In Vivo, 885-894, Copyright (2012) with permission from Elsevier. Full text @ Cell Metab.