Tagln2 regulates CV pruning by promoting EC nucleus migration.(A) Role of tagln2 in CV pruning in zebrafish embryos. Boxes show enlarged images of the CV. The arrowhead indicates unpruned vessel. Scale bar: 50 μm. (B) Quantification of vascular loops in sibling and tagln2-/- embryos: sibling, n = 25 embryos; tagln2-/-, n = 23 embryos. P = 0.0004. Student’s unpaired two-tailed t test. (C) Time-lapse live imaging of Tg(fli1a:EGFP;kdrl:mCherry) embryos shows EC nucleus migration in CV pruning in control or tagln2 morphant. Arrows indicate the direction of the blood flow. Colored circles indicate the EC nuclei. Scale bar: 10 μm. (D) Direction of EC nucleus migration in the regressing vessel. The direction is classified into three types: with the flow, static, and against the flow. A total of 12 vascular loops with 24 EC nuclei and 8 vascular loops with 15 EC nuclei were calculated in the control group and tagln2 morphant, respectively. Unpaired two-tailed chi-square test. P = 0.0471. (E) and (F) Efficiency of siTAGLN2 in HUVECs at the mRNA (P = 0.0094) and protein (P < 0.0001) levels. (G) Wound healing of siCTR-transfected and siTAGLN2-transfected HUVECs. Statistics for wound healing after siCTR and siTAGLN2 transfection: 6 h, P = 0.0037; 12 h, P = 0.0252. (E)-(G) Student’s unpaired two-tailed t test. *P < 0.05, **P < 0.01, ***P < 0.001.
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