Larger Image Figure S3. ATP4a Acts on Noncanonical Wnt Signaling, Related to Figure 4(A and B) Convergent extension. (A) Neural tube closure: widening of the neural tube in ATP4a morphants. Embryos were injected unilaterally into the animal right blastomeres at the 4 cell stage, fixed and processed for Sox3 expression to visualize the neural plate by WMISH at stage 18. Staging was according to the progress of neural tube closure on the uninjected (left) side. Note that the widening of the neural plate was partially rescued upon ATP4a co-injection. (B) Shortening of the anterior-posterior (AP) axis. Embryos were bilaterally injected into the DMZ at the 4 cell stage and the AP extension was determined at stage 32. Note that MO-induced shortening was partially rescued by ATP4a co-injection. The width of the neural plate (A) and the length of embryos (B) were measured (as indicated by dashed lines). Results are depicted as box plots.(C) Neural crest cell specification and migration: right-sided ATP4aMO-injection resulted in reduced and altered Twist1 mRNA expression at stage 26 (n1-n3/4; neural crest migratory streams).(D) Right-sided pronephric tubule (pnt) defects upon unilaterally right-sided ATP4aMO-injection, as demonstrated by WMISH using the pronephros marker gene ATP1b1. d, dorsal; l, left; pnd, pronephric duct; pnt, pronephric tubule; r, right; v, ventral. ∗∗∗, very highly significant. Numbers in brackets represent number of analyzed specimens. Image published in: Walentek P et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue sox3.S Sry, Xlsox3, xSox3, Xtsox3 X. laevis Throughout NF stage 18 neural tube chordal neural plate pre-chordal neural plate neural plate anterior neural fold neural fold twist1.L twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 26 cranial neural crest migratory neural crest cell atp1b1.S atp1b1a, Beta1-Na/K-ATPase, Na+-K+-ATPase, β1-NaK-ATPase X. laevis Throughout NF stage 32 pronephric nephron pronephric kidney pronephric tubule pronephric duct eye post-anal gut Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + atp4a MO + NF18 (in situ hybridization) fig.S3.a Anatomical Phenotype increased width of neural plate Expression Phenotype increased amount sox3.L expression in neural plate Xla Wt + atp4a MO + NF32 (morphology) fig.S3.b Anatomical Phenotype decreased length of anterior-posterior axis Xla Wt + atp4a MO + NF26 (in situ hybridization) fig.S3.c Anatomical Phenotype abnormal cell migration in neural crest abnormal development of neural crest Expression Phenotype decreased amount twist1.L expression in neural crest mislocalised twist1.L expression in neural crest Xla Wt + atp4a MO + NF32 (in situ hybridization) fig.S3.d Anatomical Phenotype absent pronephric tubule Permanent Image Page Printer Friendly View XB-IMG-82876