Comparison of junctions formed by Dn-αCat or its mutants in 468 cells. The cells were double stained for occludin to reveal TJs (top row) and for Dendra2 to reveal transgene products (bottom row). The maps of Dn-αCat proteins are shown as in Fig. 3. The mutated domains are in yellow. Dn-αCat can interact with actin both directly through αABD and indirectly through vinculin, whereas its Dn-αCat-ΔVin and Dn-αCat(1–505) mutants can only associate with actin specifically through αABD or vinculin, respectively. Both modes of interactions are inactivated in the double mutant Dn-αCat-ΔVin+I792A. Note that cells expressing intact α-catenin were the only ones to produce fully closed rings of TJs. All the mutants showed comparable levels of expression (Fig. S4 a). Bars, 40 µm.