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Figure 7 | Cilia

Figure 7

From: Multiple essential roles for primary cilia in heart development

Figure 7

Shh expression is normal but downstream Shh signaling is compromised in cbs mutants. (A-C) WISH upon E10.25 (A,B) and E11.5 (C) wild type (+/+) and cbs/cbs embryos. Arrows indicate Shh (A), Gli1 (B), and Ptch1 (C) expression in pharyngeal mesoderm adjacent to the heart tube. (D) In situ hybridization on transverse sections of E10.5 wild type (+/+) and cbs/cbs embryos, using Pitx2c as a probe. Arrowheads indicate expression in the left atrium, while the arrow indicates expression in the right atrium of cbs/cbs heart. (E) Immunofluorescence analysis of apoptosis in pharyngeal arches of E10.5 wild type (+/+) and cbs/cbs embryos using an antibody recognizing cleaved, activated caspase-3 (casp3, yellow). Blue: DAPI-labeled nuclei. (F) Individual cilia in E12.5 wild type (+/+) and cbs/cbs endocardial cushions, labelled with antibodies recognizing Arl13b (red) to mark the ciliary axoneme and γ-tubulin (green) to mark the basal bodies. Traces of DAPI-stained nuclei can be seen in dark blue. Scale bars: A, 400 μm; B,C, 500 μm; D, 200 μm; E, 20 μm; F, 1 μm. cbs, cobblestone; DAPI, 4’-6-diamidino-2-phenylindole; E10.5,embryonic day 10.5; Shh, Sonic hedgehog; WISH, whole mount in situ hybridization.

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