Fig. 7. Interaction with Ca²⁺/calmodulin is required for the translocation of l-caldesmon to podosomes. A7r5 cells transfected with plasmids encoding EGFP-CadCamAB (green) were either stained with TRITC-phalloidin to identify F-actin (red) (A,B,D,E) or with a monoclonal antibody raised against vinculin (red) to identify focal adhesions (C). The two channels were merged (overlay) to show co-localisation. (A-C) Expression of EGFP-CadCamAB caused severe disruption of actin stress fibres (A) and focal adhesions (C) in some cells; others retain an intact array of stress fibres (B). (D) Confocal micrograph of CadCamAB-transfected cells stimulated with 1 µM PDBu for 30 minutes prior to staining. Insets in D show a 10x10 µm enlargement of the cell to show the exclusion of CadCamAB from the actin core of podosomes. (E) Reconstruction of the x-z profiles of selected podosomes of the cell in D. Bars, 20 µm in A-D; 2 µm in E.