Xenbase Image ID: 154017

Figure 1. Depletion of Inner-Ring Nups Alters LR Patterning (A) One- (left) and 2-cell (right) injection schematic (not to scale) showing a salmon-colored dye as a tracer. When injections are done in one cell at the 2-cell stage, embryos are selected where either the left or the right side of the embryo is targeted, as shown in the diagram of dissected LRO (stage 16) or in a later stage-27 embryo. Small red lines in LRO represent cilia. (B) Percentage of embryos with abnormal cardiac looping (see C for key to score cardiac looping) after MO injection at 1-cell stage (left) or 2-cell stage (right). nup188 morphant heart looping defects are rescued by human NUP188 expression (left). n represents total number of embryos from three independent experiments. ∗∗∗p < 0.0005, ∗∗p < 0.005, chi-squared/Fisher's exact test. UC, uninjected controls. (C) Examples of cardiac looping phenotypes of wild-type and abnormal Xenopus hearts at stage 45 viewed ventrally with anterior at the top. D-Loop: normal outflow track (OT) loops right (short dashed line). L-Loop: abnormal OT loops left. A-Loop: abnormal OT is unlooped. Ventricle (V) is bound by a long dashed line. (D) Percentage of embryos with abnormal pitx2c (see E for key to score pitx2c staining) after MO injection at 1-cell stage (left, middle) or 2-cell stage (right). n represents total number of embryos from three independent experiments. ∗∗∗p < 0.0005, ∗p < 0.05, chi-squared/Fisher's exact test. (E) Example key of pitx2c expression showing lateral views of embryos (stages 28–31) with dorsal at the top. Arrowheads point to pitx2c in the lateral plate mesoderm. (F) Percentage of embryos with abnormal coco expression (see G for key for coco scoring) after MO injection at 1-cell stage (left, middle) or 2-cell stage (right). L and R denote left and right side of LRO, respectively. n represents total number of embryos from three independent experiments. ∗∗∗p < 0.0005, ∗∗p < 0.005, chi-squared/Fisher's exact test. (G) Example key of coco expression patterns in the LRO at stages 19–20. Ventral views with anterior at the top. (H) Schematic of NPC. Nups referred to in this study are in bold. (I) Nup188 and Nup93 protein levels determined by western blot. Numbers are relative protein levels normalized to Gapdh with triangle reflecting dose of MO. Boxed lane indicates dose used for all experiments. See also Figures S1 and S2. Image published in: Del Viso F et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue pitx2 pitx2-A, pitx2-a, pitx2-b, pitx2c, xpitx2, xPtx2 X. tropicalis Sometime during NF stage 28 to NF stage 31 lateral plate mesoderm left cement gland heart trigeminal ganglion profundus ganglion dand5 51-B6, cer2, cerl2, charon, cktsf1b3, coco, crl2, dante, grem3, S10-51-B6, xCoco X. tropicalis Sometime during NF stage 19 to NF stage 20 gastrocoel roof plate Image source: Published Experiment + Assay Source Phenotypes and Disease Xtr Wt + nup188 MO + NF45 (whole-mount microscopy) Fig 1. B C Anatomical Phenotype abnormal heart looping abnormal heart morphology Disease congenital heart disease visceral heterotaxy Xtr Wt + nup188 MO + NF45 (whole-mount microscopy) Fig. 1 B C Anatomical Phenotype abnormal heart looping abnormal heart morphology Disease congenital heart disease visceral heterotaxy Xtr Wt + nup188 MO + NF45 (whole-mount microscopy) Fig. 1 B C Anatomical Phenotype abnormal heart looping abnormal heart morphology Disease congenital heart disease visceral heterotaxy Xtr Wt + nup93 MO + NF45 (whole-mount microscopy) Fig. 1 B C Anatomical Phenotype abnormal heart looping abnormal heart morphology Disease congenital heart disease visceral heterotaxy Xtr Wt + nup188 MO + NF28-31 (in situ hybridization) Fig. 1 D E Disease congenital heart disease visceral heterotaxy Expression Phenotype absent pitx2 expression in lateral plate mesoderm mislocalised pitx2 expression in lateral plate mesoderm Xtr Wt + nup188 MO + NF28-31 (in situ hybridization) Fig. 1 D E Disease congenital heart disease visceral heterotaxy Expression Phenotype absent pitx2 expression in lateral plate mesoderm mislocalised pitx2 expression in lateral plate mesoderm Xtr Wt + nup188 MO + NF28-31 (in situ hybridization) Fig. 1 D E Disease congenital heart disease visceral heterotaxy Expression Phenotype absent pitx2 expression in lateral plate mesoderm mislocalised pitx2 expression in lateral plate mesoderm Xtr Wt + nup93 MO + NF28-31 (in situ hybridization) Fig. 1 D E Disease congenital heart disease visceral heterotaxy Expression Phenotype absent pitx2 expression in lateral plate mesoderm mislocalised pitx2 expression in lateral plate mesoderm Xtr Wt + nup188 MO + NF19-20 (in situ hybridization) Fig. 1 F G Disease congenital heart disease visceral heterotaxy Expression Phenotype mislocalised dand5 expression in left-right organizer Xtr Wt + nup93 MO + NF19-20 (in situ hybridization) Fig. 1 F G Disease congenital heart disease visceral heterotaxy Expression Phenotype mislocalised dand5 expression in left-right organizer Xtr Wt + nup188 MO + NF19-20 (in situ hybridization) Fig. 1 F H Disease congenital heart disease visceral heterotaxy Expression Phenotype mislocalised dand5 expression in left-right organizer Xtr Wt + nup93 MO + NF19-20 (in situ hybridization) Fig. 1 F H Disease congenital heart disease visceral heterotaxy Expression Phenotype mislocalised dand5 expression in left-right organizer Larger Image Printer Friendly View

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