Interaction of migfilin with kindlin-1. A, GST-kindlin-1 specifically binds to migfilin in human 293 cells. GST-kindlin-1-bound glutathione-Sepharose or control beads were incubated with cell lysates and subjected to Western blotting using anti-migfilin antibody. Lane 1, cell lysate; lane 2, cell lysate incubated with GST-Sepaharose beads; lane 3, cell lysate incubated with GST-kindlin-1 beads. B, Immunolocalization of kindlin-1 (green) at the end of the actin filaments (red) in keratinocytes; C, immunolocalization of migfilin (red) at the end of actin filaments in keratinocytes; D–F, double immunolocalization of kindlin-1 (D, green) and migfilin (E, red) in same focal adhesions (F, orange). Arrows point to the focal adhesions.