Figure 5

Endothelial Rudhira is essential for vascular patterning. Control and rudh^CKO at E10.5 and E11.5 were analyzed as indicated. (a) Unstained embryos, (b,c) Histological analysis showing comparison of heart and dorsal aorta (DA). (d) Immunostaining analysis of heart, dorsal aorta (DA) and intersomitic vessels (ISVs) using myocardial marker SMA, primitive erythroid marker Ly76 and vascular marker PECAM. Nuclei are marked by DAPI (Blue). Black or white dotted lines mark the boundary of DA. Graph shows quantitation of dorsal aorta width in the thoracic region. (e) Whole mount PECAM staining in control and rudh^CKO embryos as indicated, with magnified view of head and intersomitic vessels. Arrows: normal vascular patterning; arrowheads: irregular and discontinuous vasculature. N = 5 embryos. Results shown are a representative of at least three independent experiments with at least three biological replicates. Statistical analysis was carried out using one-way ANOVA. Scale bar: (a) 500 μm; (b,c) 100 μm; (d) E10.5 and E11.5 heart: 200 μm, E10.5 and E11.5 DA: 50 μm, ISV: 50 μm; (e) 500 μm. *p < 0.05, **p < 0.01, ***p < 0.001. The observed chi-square values for two degrees of freedom were 0.88 (E9.5), 2.62 (E11.5), 2.62 (E12.5), 25.68 (E13.5) and 37.5 (Postnatal).