Kindlin-2 knockdown and focal adhesion localization. A. Confocal immunofluorescent microscopy with anti-β1 integrin (green) and anti-paxillin (red) on C2C12 cells transfected with RNAi and then changed to differentiation media for 2 days. Control cells (scr RNAi) show linear staining consistent with localization to costameres (arrows), as well as punctate focal contact staining (arrowheads). Conversely, focal contact proteins in the kindlin-2 RNAi cells fail to form linear structures and instead are concentrated in unusual appearing puncta (*). (Scale bar = 20 μM). B. Oregon Green phalloidin staining on day 2 differentiated C2C12 cells. In the scrambled RNAi myocytes, actin is found primarily in a linear, fiber like pattern. Conversely, in the kindlin-2 RNAi cells it is found concentrated at the membrane. (Scale bar = 20 μM).