XB-IMG-82990
Xenbase Image ID: 82990
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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-3xCâ²egfp 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.
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