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. Author manuscript; available in PMC: 2011 Jul 1.
Published in final edited form as: Nat Cell Biol. 2010 Dec 19;13(1):49–58. doi: 10.1038/ncb2133

Figure 1. DDR1 is required for collective cell migration.

Figure 1

(a) Top-left panel shows A431 SCC cells collectively invading a 3D matrix (F-actin – red, reflectance – cyan). Other left-hand panels show higher magnification of the indicated area. β-catenin – blue, F-actin – red, GFP-MLC – green. Right-hand panels show A431 cells on a 2D substrate, colours as in left-hand panels. Scale bars are 10 μm. (b) DDR1 western blots showing efficacy of siRNA (upper panel) and shRNA (lower panel); tubulin is used as loading control. (c) Representative images of F-actin organisation, pS19-MLC, myosin IIa or E-cadherin in A431 cells transfected with control siRNA (left) or with two different siRNA oligonucleotides against DDR1: #3 (middle) and #4 (right). Scale bars are 20 μm. (d) i) Upper panels show snap shots from phase contrast movies of control and DDR1 shRNA-transfected A431 cells moving on collagen gels. Scale bars are 20μm. Blue shading shows the position of cell groups at t=0 and red shading shows the position of the same cell groups at t=6 hours. ii) and iii) show the cell dispersion index and the average speed of cells within groups, respectively. Data calculated from tracking multiple cell groups from multiple experiments (analysis of 7-10 colonies from three independent experiments). * indicates p<0.01 student’s t-test. (e) Panels show 3D reconstruction of ‘spheroid’ SCC invasion assays in control and DDR1 depleted A431 cells. Grid spacing is 50μm. (f) Panels show H&E sections of control or DDR1 siRNA transfected SCC cells in an organotypic assay. The number in the bottom-left corner shows the invasion index of three independent experiments as percentage of the non-transfected cells.