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. 2012 Dec 24;199(7):1103–1115. doi: 10.1083/jcb.201207009

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© 2012 Ngok et al.

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Figure 2. — Cell junction and heart defects in the syx^−/− mouse. (A) Confocal images of confluent monolayers of ZO1-immunolabeled (green) syx^+/+ and syx^−/− ECs, acquired with the same imaging settings. Arrows point to appositions between syx^−/− ECs where ZO1 is absent. The framed regions are magnified in the insets. Bar, 50 µm. (B) Quantification of the percentage of ECs surrounded by ZO1 in fields of syx^+/+ or syx^−/− ECs, each containing ∼65 cells (n = 10; ±SEM; *, P < 1.0⁻¹⁰). (C) Expression levels of ZO1 in syx^+/+ and syx^−/− ECs. The β-actin immunoblot is a gel loading control of the ZO1 lanes. (D) syx^+/+ and syx^−/− ECs were plated as confluent monolayers. Impedance at 4,000 Hz was measured every 180 s for 48 h (means ± SEM). (E) Images of syx^+/+, syx^−/−, syx^fl/fl, and tie2-Cresyx^fl/fl lectin-stained (red) tracheal venules injected with fluorescent microspheres (green, appearing yellow because of overlap with vessel walls). Arrows point to extravasated microspheres along the walls of syx^−/− vessels. Bar, 25 µm. (F) Relative values of extravasated bead fluorescence in images of tracheal vessels (n = 6; ±SEM; *, P < 0.009 and 0.0008). (G) Transmission electron micrographs of thin sections of syx^+/+ and syx^−/− left ventricle myocardium. The junctions between ECs of syx^+/+ capillaries are sealed, but EC junctions of syx^−/− capillaries are open or malformed. The framed regions are magnified below. Bars, 1 µm. (H) Images acquired as in G show that erythrocytes are enclosed within capillaries in the syx^+/+ myocardium but are present in the interstitial space of the syx^−/− myocardium. Bars, 5 µm. (I) Representative series of pressure–volume loops measured in syx^+/+ and syx^−/− left ventricles. Dashed lines denote end-diastolic pressure–volume relation (EDPVR) approximations, whose slopes (0.38 and 0.74 mmHg/µl in the syx^+/+ and syx^−/− ventricles, respectively) represent the passive stiffness of the ventricular wall. The data shown are from a single representative experiment out of three repeats. (J) syx^+/+ and syx^−/− EDPVR slopes (means ± SD; n = 3; *, P = 0.015) normalized by the lowest slope of the syx^+/+ EDPVR.