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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. |
