Fig. 1. CMS colocalizes with F-actin to dynamic actin structures such as membrane ruffles and podosomes. NIH 3T3 cells, COS-7 cells and podocytes transfected with CMS, were subjected to immunofluorescence. (A) Endogenous paxillin and transiently expressed FLAG-tagged CMS in NIH 3T3 cells showed a different cellular localization pattern, indicating that the cytoplasmic CMS protein is concentrated at leading edges of migratory cells and not to focal adhesions as seen for paxillin. Bars, 20 µm. (B) COS-7 cells transiently expressing CMS were stimulated with PMA for 10 minutes, stained for CMS and F-actin and analyzed by confocal microscopy. CMS was concentrated and colocalized in prominent membrane ruffles indicated by arrows. Bars, 20 µm. (C) Mouse podocytes stably overexpressing Myc-tagged CMS were differentiated for 7 days and subsequently stained for CMS and F-actin. CMS colocalizes with dynamic actin structures such as lamellipodia and podosomes (see arrows). Bars, 10 µm.