XB-IMG-171225
Xenbase Image ID: 171225
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Figure S1. Related to Fig. 1; Depletion of Rapgef5. (A) Knockdown of Rapgef5
disrupts cardiac looping. Ventral views of Xenopus tadpoles with anterior to the top.
Representative examples of a normal (D-loop outlined in blue lower panel), and
abnormal (A-Loop outlined in green, L-Loop outlined in red) looped heart. Injection of
a control MO or a control CRISPR (targeting tyrosinase) does not affect cardiac
looping. (B) Injection of a control MO or control CRISPR (targeting tyrosinase) does
not cause abnormalities in pitx2c expression. (C) The control (tyrosinase) CRISPR
reduces pigment levels in stage 45 Xenopus embryos, demonstrating that the
CRISPR/CAS9 system functions as expected. (D) Rationale of R5 MOSplice design. The
splice MO targets the splice donor site at the start of intron 2. Failure to excise intron
2 results in a premature stop codon early in the Rapgef5 protein. RT-PCR confirms
that R5 MOSplice causes retention of intron 2. In genomic DNA, intron 2 spanning
primers amplify a band of 547bp. In WT cDNA, these primers amplify a band of 157bp.
PCR primers amplifying cDNA from R5 MOSplice morphants generate a band at 547bp
indicating retention of intron 2 in mRNA. A “no template” negative control is also
included. A single asterisk indicates P<0.05 and a triple asterisk P<0.005. Image published in: Griffin JN et al. (2018) Copyright © 2018. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |