Xenbase Image ID: 156484

Fig. 4. PlexinA1 and PTK7 interact in immunoprecipitation and phenotypic assays. (A) Full length HA-tagged PlexinA1 (PlxA1-HA) precipitates myc-tagged PTK7 (PTK7-myc). Upper panels show the immunoprecipitated proteins, the lower panels the Xenopus lysates. Embryos were injected with 800 pg PTK7-myc RNA and 1 ng PlexA1-HA RNA. 50 Embryos were lysed at gastrula stage and used for immunoprecipitation. Injected constructs and antibodies used for Western blotting are indicated. (B–F) Co-expression of PlexinA1 and PTK7 leads to an increase in NC migration defects. Xenopus embryos were injected with the respective RNA construct in combination with 75 pg lacZ RNA in one blastomere at the two-cell stage. RFP RNA was co-injected to equalize injected RNA concentrations. NC migration was analyzed by twist in situ hybridization at neurula stage 22. (B) Graph summarizing the percentage of NC migration defects of three independent injection experiments. Number of injected embryos is indicated for each column. (∗) Percentage of NC migration defects caused by co-injection of PlxA1 and PTK7 is significantly different from the PlxA1 injected embryos in a student t-test (p < 0.05). (C–F) Twist in situ hybridization of injected neurula stage 20–22 embryos, the injected lacZ-positive (blue) side is shown on the right. (C) Embryo injected with 1 ng RFP RNA shows normal NC migration. (D) Embryo injected with 500 pg PTK7 RNA and 500 pg RFP RNA showing normal NC migration. (E) Embryo injected with 500 pg PlexinA1 RNA and 500 pg RFP RNA. NC migration is affected (white arrow). (F) NC migration is inhibited in an embryo co-injected with 500 pg PlexinA1 and 500 pg PTK7 RNA. NC cells are induced, but fail to migrate (white arrow) (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.). Image published in: Wagner G et al. (2010) Copyright © 2010. This image is 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 twist1.L twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 20 cranial neural crest Image source: Published Larger Image Printer Friendly View

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