Morpholinos against ptges and ep4 transcripts are effective and demonstrate synergism. (A) RT-PCR analyses of ptges transcripts from total RNA isolated from 15 embryos in uninjected control (ctrl), 2ng of ptges-MO2 injected, and 4ng of ptges-MO2 injected embryos. Diagram demonstrates the loss of exon2 in ptges-MO2 injected embryos due to blocking of splicing acceptor site. This results in alternative splicing from the splicing donor site at the end of exon1 and acceptor site in the beginning of exon3. This was verified by sequencing of the 428bp band. (B) Images of embryos overexpressing GFP-tagged EP4 receptor injected with ep4-gfp RNA alone or co-injected with ep4-MO2. (C) ep4- and ptges-MO reveal synergism at sub-optimal doses. Injection of ep4- or ptges-MO at 1ng cause mild shortening of zebrafish length at 24 hpf. However co-injection of ep4- and ptges-MO at 1ng results in synergistic effect on the length of zebrafish embryos at 24 hpf.
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