XB-IMG-171780
Xenbase Image ID: 171780
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Figure S1. Generation of the Double Heterozygous Line for Brachyury
paralogues t and t2, Related to Figure 1
(A) Scheme to generate the t
e1.2D/+t2e3.7D/+ (t
-/+t2-/+) X. tropicalis line. (B,F) TALEN
design for t or t2 mutagenesis and positions of MOs blocking donor splice site (MOsplice)
or translation initiation site (MOtransl) of the corresponding transcript. (C,G) TALENinduced
mutation rate at the targeted SacI or EcoRI site as estimated by the partial
restriction digest of specific PCR amplicons. (D) Sanger sequencing summary of
generated indels in exon 1 of t. (E) Morphological defects caused by TALEN-induced
t mutations at late tailbud stage. Scale bar, 0.5 mm. (H) Western blot of injected wildtype
and mutant t or t2 constructs tagged either N- or C-terminally with HA. The
detection of exogenous myc (as part of the injected fam83g-myc mRNA) and
endogenous α-tubulin were used as controls for injection/translation efficiency and gel
electrophoresis loading, respectively. (I) Mutant Brachyury constructs failed to disrupt
gastrulation. Scale bar, 0.25 mm. Image published in: Gentsch GE et al. (2018) © 2018 The Francis Crick Institute. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |