Xenbase Image ID: 83945

Fig. 9. Trio interacts with Par3 and TrioMO rescues Par3 morphants in Xenopus. (A) Co-immunoprecipitation of Trio and Par3 in cells expressing TrioHA and Par3GFP. Following pull-down with the HA tag, western blots against HA showed a band at 320 kDa corresponding to TrioHA and western blots against GFP showed a band at 250 kDa corresponding to Par3GFP. Controls using no transfected cells or control IgG antibody showed no HA or GFP staining. Input lanes show that TrioHA and Par3GFP were present and ran at 320 kDa and 250 kDa, respectively. (B-E) Colocalisation analysis in Xenopus NC cells. Par3RFP (B) and TrioGFP (C) are both found at cell-cell contacts, where they colocalise (D). (E) Magnification and colocalisation mask area indicated in D. Scale bar: 20 μm. (F) Dispersion analysis in control (black), Par3MO (red) or Par3Mo/TrioMO (white). n=10 explants for each condition, more than 30 cells analysed per explant; *P<0.05, **P<0.01, ***P<0.001. (G) Speed of migration. Par3MO, n=70; P=0.4642; Par3MO+TrioMO, n=90; P=0.2713. (H) Persistence. Par3MO, n=70; P=0.4637; Par3MO+TrioMO, n=90; P=0.1126. (I-K) Cell collision assay. CIL was analysed with the cell collision assay previously described. (I) Normal CIL behaviour in control cells. n=10; P<0.001. Red arrows indicate the initial velocity vector. (L-P) Twist expression in control embryos (L) or those injected with Par3MO (M) or Par3MO+TrioMO (N). Asterisk indicates the eye. (O) Percentage of embryos with normal NC migration. Control, n=9; Par3MO, n=13; **P<0.01; Par3MO+TrioMO, n=18; P<0.05. (P) Distance migrated by NC cells relative to mean migration in control embryos. Migration is reduced in Par3MO-injected embryos (***P<0.001) but co-injection of TrioMO with Par3MO increases migration (***P<0.001). Error bars indicate s.d. Image published in: Moore R et al. (2013) Copyright © 2013. 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 twist1.S twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 24 neural crest cranial neural crest mandibular crest hyoid crest Image source: Published Larger Image Printer Friendly View

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