α-Catenin and α-actinin do not colocalize. (A) Immunostaining in WT MEFs for α-actinin shows its colocalization after 15 min of spreading with F-actin at the cell edge (yellow arrow) as well as its localization in nascent adhesions (white arrow). (B) Zoom-in on the edge of a WT MEF cell costained for actin, α-actinin, α-catenin, showing lack of overlap of the latter two. (C) Immunostaining for α-actinin and α-catenin in WT MEFs shows their lack of overlap in actin stress fibers. Bottom row is the zoom-in of the boxes in the top row; arrows point to locations in which α-catenin is localized but α-actinin is not. (D) Frame from a video of a cell expressing GFP-α-catenin and mCherry-α-actinin showing that the two are not colocalized; and kymographs taken from the yellow line (D′).