RhoC negatively regulates VEGF-induced vascular permeability in zebrafish. (A,B) Using zebrafish cDNA, a probe to zebrafish RhoC (Rhoad) was created and in situ hybridization was performed on 24hpf zebrafish embryos. Multiple images were captured using a Zeiss Axioplan 2 microscope and overlayed in Photoshop, such that areas of focus were unmasked, to generate a composite image. Lateral (A) and superior (B) views are shown. DA, dorsal aorta; PCV, posterior cardinal vein; ISVs, anterior intersomitic vessels; NT, neural tubes. (C) Microangiography was performed on anesthetized 3dpf zebrafish embryos by injecting FITC–dextran (2000kDa) and Texas-Red–dextran (70kDa), VEGF was induced through heat exposure (when applicable), and extravasation of red tracer as a measure of zebrafish vascular permeability was live imaged using a ZEISS LSM 780 confocal microscope. Control, no MO injection and no VEGF induction; Cont MO, control MO injection; RhoC MO, Rhoad and Rhoae MO injection; Unind, no VEGF induction; VEGF, heat induction of VEGF transgene. (D) Quantification of extravasated red tracer. **P<0.05 (RhoC MO, VEGF Induced versus Control MO, VEGF Induced); *P<0.05 (RhoC MO, Uninduced versus Control; Control MO, VEGF Induced versus Control) (paired two-tailed Student′s t-test). Results are mean±s.d. (n=??).