Xenbase Image ID: 153874

Fig. 4. Lhx9 regulates integrin α4-paxillin signaling in PE clusters. (A-D) Whole embryo in situ hybridization for itga4, stages 36 to 46. Lateral views of anterior portion (left panels) with magnified image of cardiac region (right panels). Red arrowheads indicate PE cluster; asterisks indicates heart. (E,F) In situ hybridization for itga4 in control (E) and Lhx9-depleted embryos (F) at stage 41. Representative cardiac region shown, anterior to right, dorsal to top; red arrowhead marks PE cluster. (O) Quantification of observed itga4 expression (reduced expression denoting ≤50% stain intensity compared with controls) and clustering defects shown in E,F (see Fig. 2M for phenotype assessment), embryos taken from six independent experiments, P≤0.0001 by Chi-square test. (G-J′) Transverse cardiac region agarose sections from control (G-H′) and Lhx9-depleted (I-J′) stage 41 embryos post-in situ hybridization for tcf21 (G,I), nuclei expression with DAPI (H,J, blue) and Itgβ1 immunohistochemistry (H′,J′, magenta). White outlines (H,H′,J,J′) indicate PE cluster as depicted by tcf21 and Itgβ1. (K-N′) Transverse cardiac region agarose sections from control (K-L′) and Lhx9-depleted (M-N′) stage 45 Xla.Tg(Cardiac-actin:GFP)Mohun embryos depicting representative immunohistochemical analysis for DAPI (blue), GFP (green) to label cardiomyocytes, Itgβ1 (magenta) to label epicardial cells and phosphorylated Y118-paxillin (red). Magnified images (L,L′,N,N′) from white boxes in K,M. White arrowheads indicate PE cluster. (P) Pixel intensity (integrated density) levels for phosphorylated Y118-paxillin from five control and 10 Lhx9-depleted embryos; P=0.0134 by two-tailed Student’s t-test. v, ventricle. Image published in: Tandon P et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue itga4.L inta4 X. laevis Throughout NF stage 39 proepicardium itgb1.L Beta 1 integrin, cd29, fnrb, gpiia, itgb1-a, itgb1-b, mdf2, msk12, vlab, vla-beta X. laevis Throughout NF stage 41 proepicardium itga4.L inta4 X. laevis Throughout NF stage 41 proepicardium tcf21.L capsulin, epicardin, pod 1, pod1 X. laevis Throughout NF stage 41 proepicardium itgb1.L Beta 1 integrin, cd29, fnrb, gpiia, itgb1-a, itgb1-b, mdf2, msk12, vlab, vla-beta X. laevis Throughout NF stage 45 epicardium heart pxn.L paxillin X. laevis Throughout NF stage 45 heart epicardium itga4.L inta4 X. laevis Throughout NF stage 46 epicardium Image source: Published Larger Image Printer Friendly View

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