Xenbase Image ID: 129882

Fig. 2. Rab3d is required to negatively regulate BMP signaling during anterior neurulation. A–D: Rab3d MO (20 ng), dnSmad1GR (1 ng), and Smad1GR (500 pg) were co-injected with a lineage tracer b-galactosidase mRNA as indicated. Dexamethasone was applied to the culture media to activated GRfused protein products. Insets show the same embryos with low magnification. Data are the representative of three independent experiments. A,B: At stage 19, embryos were subjected to in situ hybridization with Sox2 probe. Arrows indicate the injected sides. Dashed lines mark the midlines of the embryos. (Sox2 down: reduced sox2 expression without CV defect; CV defect: convergence defect). C,D: At stage 19, neural plate folding defects were counted. Arrows indicate typical pigmentation of neural folds. E: Rab3d MO increased phosphorylated Smad1 level. Rab3d MO and Rab3d mRNA were co-injected into dorsal-animal blastomeres of 4-cell-stage embryos. At stage 19, embryos were collected, lysed, and subjected to western blot with a- P-Smad1/5 (Cell Signaling, Danvers, MA), a-Smad1 (Santa Cruz Biotechnology), or a-actin (Santa Cruz Biotechnology). Data are representative of three independent experiments. F: BMP responsive reporter (SBE4-Luc) activities were measured from control and Rab3d MO (10 ng) injected embryos. SBE4-Luc, along with pRLTk (renilla luciferase as an internal standard), were injected with or without Rab3d MO as indicated. At neurula stage, embryos were separated into four or five pools of five embryos for assay in multiplicate and subjected to the dual luciferase assay. Image published in: Kim H and Han JK (2011) Copyright © 2011. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue sox2.L anop3, LOC108718087, mcops3, Sox-2, XLSOX-2, Xsox-2, XSox2 X. laevis Throughout NF stage 19 neural plate Image source: Published Larger Image Printer Friendly View

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