Retardation of junction formation by Apg-2 depletion. Control and Apg-2 RNAi cell lines were cultured and processed as in figure 3 and then stained with antibodies against the transmembrane proteins occludin (A) and claudin-4 (B), the junctional plaque components cingulin (C) and GEF-H1 (D), and the adherens junction proteins α-catenin (E) and E-cadherin (F). Shown are images derived from samples fixed after 1 hour and 27 hours of junction formation. Note, retardation of junctional recruitment was observed for occludin, cingulin, α-catenin and E-cadherin. GEF-H1 became only concentrated at tight junctions after longer time points of junction formation. Claudin-4 was present at the plasma membrane at all time points but only appeared to be concentrated at junctions at later time points. GEF-H1 and claudin-4 distributions were not affected by depletion of Apg-2.