Xenbase Image ID: 82990

Fig. 3. Itpkb accumulates around the wound edge and modulates F-actin assembly in multicellular wounds. (A) Immunofluorescence of wounded embryos shows the accumulation of endogenous Itpkb at the wound edge. Embryos were injected with membrane GFP probe (gap-43-gfp mRNA), wounded, fixed, and stained with antibodies against GFP and Itpkb. Green, GFP; red, Itpkb. n = 6. (B) Itpkb accumulates at the leading edge of wounded cells. (Left) An animal cap wound in an embryo injected with 0.2 ng egfp mRNA as a control. (Right) An animal cap wound in an embryo injected with 5 ng itpkb-3xC′egfp mRNA (itpkb-eGFP). Control n = 7, itpkb-eGFP n = 8. (C) Itpkb colocalizes with the F-actin array around the wound edge. Embryos were coinjected with 1.5 ng cherry-dbmoesin mRNA and 5 ng itpkb-egfp mRNA. Green, itpkb-eGFP accumulation around wound edge; red, F-actin array around the multicellular wound; yellow, merged image of F-actin and itpkb-eGFP postwounding (p.w.). The image represents an example at 2 min p.w. n = 7. (D) The accumulation of Itpkb at the leading edge of wounded cells is F-actin�dependent. Embryos were injected with itpkb-egfp mRNA, treated with DMSO (Left), LatB (2.5 μM) (Center), or Noc (50 μM) (Right) for 30 min before wounding. Live images were taken 3 min after wounding to observe the localization of itpkb-eGFP. N DMSO = 11; N LatB = 9; N Noc = 8. (E) Itpkb knock down impairs F-actin assembly after wounding. Embryos were coinjected with 0.5 ng F-actin probe and 40 ng, itpkb-mo, or moc. Itpkb-mo embryos were also coinjected with myc-itpkb mRNA. (Left) F-actin array in moc-wounded animal cap. Middle panel shows disrupted F-actin array on itpkb-mo wounded animal. (Center) Rescued F-actin array in itpkb-mo + myc-itpkb injected embryos. N moc = 14; N itpkb-mo = 20; N rescue = 9. (Bars, 50 μm.) Image published in: Soto X 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 itpkb X. tropicalis Throughout NF stage 9   Image source: Published Larger Image Printer Friendly View

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