Figure 6.
Figure 6. Inactivation of β-TRCP2 leads to VEGFR2 up-regulation and increased angiogenesis in zebrafish in vivo. (A and B) Vascular development in control (A) or 20 µM MO (B)-treated KdrI:EGFP embryos. MO was used to deplete both Fbxw11a and Fbxw11b. Images were taken by fluorescence microscopy at 72 h postfertilization (hpf). Bars, 50 µm. Data shown is representative of three independent experiments. (C and D) Close-up images of blood vessel formation in the head/anterior trunk of zebrafish treated with control (C corresponds to the blue-boxed regions of A) or Fbxw11a and Fbxw11b MOs (D corresponds to the blue-boxed regions of B). Yellow arrows in C point to junction areas where normal mirror-paired intersomitic vessel development was observed, whereas red arrows in D point to areas of excess/disorganized branching of intersomitic vessels. Bars, 50 µm. Data shown is representative of three independent experiments. (E and F) Close-up images of blood vessel formation in the posterior trunk/tail of zebrafish treated with control (E corresponds to the red-boxed regions of A) or Fbxw11a and Fbxw11b MOs (F corresponds to the red-boxed regions of B). Yellow arrows in E point to junction areas where normal mirror-paired intersomitic vessel development was observed, whereas red arrows in F point to areas of excess/disorganized branching of intersomitic vessels. Bars, 50 µm. Data shown is representative of three independent experiments. (G) RT-PCR analysis of endogenous Fbxw11a and Fbxw11b mRNA levels in MO-treated zebrafish compared with sibling controls (C). Data shown is representative of three independent experiments. (H) VEGFR2 protein expression was analyzed by Western blot in control (C) and β-TRCP2–MO-treated zebrafish embryos. Data shown is representative of two independent experiments. (I and J) Close-up images of blood vessel formation in the red-boxed region in the posterior trunk/tail of zebrafish treated with combination treatment of splice MO against Fbxw11a and Fbxw11b. Red arrows point to areas of excess/ disorganized branching of intersomitic vessels observed after depletion of both Fbxw11a and Fbxw11b. Bars, 50 µm. Data are representative of three independent experiments.

Inactivation of β-TRCP2 leads to VEGFR2 up-regulation and increased angiogenesis in zebrafish in vivo. (A and B) Vascular development in control (A) or 20 µM MO (B)-treated KdrI:EGFP embryos. MO was used to deplete both Fbxw11a and Fbxw11b. Images were taken by fluorescence microscopy at 72 h postfertilization (hpf). Bars, 50 µm. Data shown is representative of three independent experiments. (C and D) Close-up images of blood vessel formation in the head/anterior trunk of zebrafish treated with control (C corresponds to the blue-boxed regions of A) or Fbxw11a and Fbxw11b MOs (D corresponds to the blue-boxed regions of B). Yellow arrows in C point to junction areas where normal mirror-paired intersomitic vessel development was observed, whereas red arrows in D point to areas of excess/disorganized branching of intersomitic vessels. Bars, 50 µm. Data shown is representative of three independent experiments. (E and F) Close-up images of blood vessel formation in the posterior trunk/tail of zebrafish treated with control (E corresponds to the red-boxed regions of A) or Fbxw11a and Fbxw11b MOs (F corresponds to the red-boxed regions of B). Yellow arrows in E point to junction areas where normal mirror-paired intersomitic vessel development was observed, whereas red arrows in F point to areas of excess/disorganized branching of intersomitic vessels. Bars, 50 µm. Data shown is representative of three independent experiments. (G) RT-PCR analysis of endogenous Fbxw11a and Fbxw11b mRNA levels in MO-treated zebrafish compared with sibling controls (C). Data shown is representative of three independent experiments. (H) VEGFR2 protein expression was analyzed by Western blot in control (C) and β-TRCP2–MO-treated zebrafish embryos. Data shown is representative of two independent experiments. (I and J) Close-up images of blood vessel formation in the red-boxed region in the posterior trunk/tail of zebrafish treated with combination treatment of splice MO against Fbxw11a and Fbxw11b. Red arrows point to areas of excess/ disorganized branching of intersomitic vessels observed after depletion of both Fbxw11a and Fbxw11b. Bars, 50 µm. Data are representative of three independent experiments.

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