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Fig. 7. ROCK inhibition rescues the cardiac morphogenetic defects seen in SHP-2N308D embryos. (A-D) Brightfield images of stage 40 Xenopus embryos from appropriate conditions. Note that pericardial edema in SHP-2N308D embryo (3.6 ng; C) is absent in sibling incubated with Y27632 (D) and uninjected controls (A,B). Scale bar: 2 mm. (E-H) Immunofluorescent images of cleared hearts stained with anti-MHC, indicating the rescue of cardiac malformation and looping defects in SHP-2N308D embryo (G) compared with sibling incubated in Y27632 (H) and uninjected controls (E,F). Scale bar: 100 μm. (I) Graph of percentage of normal heart morphology at stage 40 in control and SHP-2N308D sibling embryos treated with 10 μM ROCK inhibitor Y27632. Statistical significance is denoted as *P≤0.0001, **P=0.0002, ***P=0.0001 using Fisher’s exact two-tailed test. Anterior is to the left, posterior to the right in all images. a, atrium; oft, outflow tract; v, ventricle. |
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Fig. S1. Associated Noonan syndrome mutations in SHP-2 lead to heart defects. Fluorescent images of anti-MHC-stained hearts from cleared stage 36 Xenopus embryos injected with SHP-2N308D (B), SHP-2D61G (C) or SHP-2Q79R (D) (3 ng) compared with uninjected controls (A). Anterior is to the left, posterior to the right. a, atrium; oft, outflow tract; v, ventricle. Scale bar: 100 . |
