FIGURE

Fig. 3

ID
ZDB-FIG-180918-5
Publication
Lisowska et al., 2018 - Cerebral Cavernous Malformation 1/2 complex controls ROCK1 and ROCK2 complementary functions for endothelial integrity
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Fig. 3

ROCK1 silencing restores normal adhesive and contractile phenotypes in CCM1-depleted HUVECs and zebrafish ccm1 mutant CCV endothelial cells. (A) Representative phase contrast images of siRNA-transfected HUVEC monolayers. (B) Representative immunofluorescence of sparsely plated siRNA-transfected HUVECs stained for F-actin (gray) and merged images with pMLC. (C) Percentage of cells with transversal actin stress fibers. Error bars, s.e.m. (n=4 except for CCM1+ROCK2 and CCM2+ROCK2 where n=2). (D) Representative immunofluorescence of F-actin (gray) and merged images with VE-cadherin of HUVEC monolayers on 20 μg/ml FN-coated coverslips at 72 h post plating. The asterisk indicates the zone of detachment for intercellular junctions. (E) Quantifications of VE-cadherin thickness from immunofluorescence staining of HUVECs presented as a percentage of control. The CT and CCM1 data are also shown in Fig. 1I. Error bars, s.e.m. (n=4 except for CCM1+ROCK1 where n=3 and CCM1+ROCK2 where n=2). In C and E, *P<0.05, **P<0.005, ***P<0.0005; NS, not significant (one-way ANOVA with Dunett's multiple comparisons). (F) Representative immunofluorescence of F-actin staining and Tg(kdrl:EGFP)-labeled endothelial cells (insets) of the CCV. (G) Quantification dot plot with s.e.m. of the intensity ratios of peripheral cortical actin over intercellular actin in WT (n=13), krit1/ccm1ty219c (16 embryos, 42 cells), krit1/ccm1ty219c+rock1-MO (16 embryos, 47 cells), and WT+rock1-MO (12 embryos, 26 cells) CCV regions; results were normalized to the WT ratio and the experiment was performed three times on independent zebrafish cohorts. ***P<0.005 (one-way ANOVA with Tukey's multiple comparisons post-test). Scale bars: 50 μm (A) 10 μm (B,D,F). CT, control.

Expression Data
Gene:
Fish:
Knockdown Reagent:
Anatomical Term:
Stage: Long-pec

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ J. Cell Sci.