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Figure 5


Fig. 5. Cx43{alpha}1KO CNCs show alteration in the actin cytoskeleton. (A,B) Rhodamine phalloidin staining showed parallel alignment of actin stress fibers in wild type CNCs (A), whereas in Cx43{alpha}1KO CNCs, stress fiber bundles were oriented in a polygonal array around the cell periphery (B). (C,D) Double immunostaining with rhodamine phalloidin (red) and vinculin (green) showed actin stress fibers typically terminated at focal adhesions in wild-type CNCs (C), but in Cx43{alpha}1KO cells (D), some actin stress fibers were not associated with focal adhesions (arrow). (E-I) Phase-contrast images show morphology of CNCs before (E) and after (F) cytochalasin D treatment, and 1 hour post-removal (G) of cytochalasin D. As CNCs re-establish their normal cell morphology, rhodamine phalloidin staining showed wild-type CNCs reformed parallel actin filament bundles (H), while Cx43{alpha}1KO CNCs reformed a multiangular ring of actin filaments around the cell periphery (I). Nuclei in A,B,H,I are delineated in blue using overlay of phase contrast images. Scale bars: 50 µm. Magnification is the same in A-D,H,I and in E-G.