Journal: Journal of the American Heart Association

Article Title: Protective Role of Endothelial Fibulin‐4 in Valvulo‐Arterial Integrity

doi: 10.1161/jaha.122.026942

Figure Lengend Snippet: Figure 7. VICs are activated in the aortic valves of DKO mice. A, Immunostaining with CD31 (known as an endothelial marker [red]), αSMA (green), and Hoechst (blue) in aortic valve leaflets from CTRL (n=5), ECKO (n=4), SMKO (n=4), and DKO (n=5) mice at postnatal day 14. High magnified images for single channel with Hoechst are shown on the right (white box). Percentage of CD31+ cells coexpressing αSMA per leaflet is shown on the right. Bars are means ± SEM. ***P<0.001, 1-way ANOVA. B–D, Immunostaining with CD31 (red) and αSMA (gray in B), CTGF (gray in C) and Gdf15 (gray in D) in aortic valve leaflets from CTRL (n=5), ECKO (n=5), SMKO (n=5), and DKO (n=5) mice at 2 months old. DAPI (blue) is shown and high magnified images are indicated in the white-dashed box. Quantification of mean signal intensity is shown on the right. Bars are means ± SEM. **P<0.01, ***P<0.001, 1-way ANOVA. αSMA indicates α-smooth muscle actin; CD31, cluster of differentiation 31; CTGF, connective tissue growth factor; CTRL, control; DKO, double knockout; ECKO, endothelial cell–specific knockout; Gdf15, growth differentiation factor-15; SMKO, smooth muscle cell–specific knockout; and VICs, valve interstitial cells.

Article Snippet: Antibody Dilution Source Catalog number CD31 (PECAM) 1:100 or 1:200 BD Biosciences #550274 αSMA 1:200 or 1:300 Sigma-Aldrich A5228 CTGF 1:200 Santa Cruz Biotechnology sc-14939 Gdf15 1:100 Santa Cruz Biotechnology sc-515675 Hoechst 1:1000 Sigma-Aldrich B2261 CD31, cluster of differentiation 31; PECAM, platelet endothelial cell adhesion molecule 1; αSMA, αsmooth muscle actin; CTGF, connective tissue growth factor; Gdf15, growth differentiation factor-15.

Techniques: Immunostaining, Marker, Control, Double Knockout, Knock-Out