Fig. 3. Detergent-resistant association of migfilin with filamentous actin at cell-cell junctions. (A-C) HaCat cells were subject to cytoskeletal extraction as described in Materials and Methods. The cells were then double stained with the mouse anti-migfilin (green, A) and rabbit anti-ß-catenin (red, C) antibodies (arrowheads). The images were merged and are shown in B. (D-F) HaCat cells were Triton X-100-extracted (D), unroofed (E), or extracted and unroofed (F) as described in Materials and Methods. The cells were then co-stained with the monoclonal anti-migfilin antibody, which was detected with an Alexa 488-conjugated anti-mouse IgG antibody (green), Rhodamine-conjugated phalloidin (red) and Hoechst dye 33258 (blue). Bars, 25 µm. (G-I) Schematic representation of the Triton X-100-extracted and/or the cationic colloidal silica unroofed cells.