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. 1999 Apr 19;145(2):347–361. doi: 10.1083/jcb.145.2.347

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Effects of α-adducin mutants on the membrane ruffling in the cells introduced with the dominant negative Rho-kinase. MBP, HA-α-adducin (WT), HA-α-adducin-AA, or HA-α-adducin-DD (10 mg/ml) was comicroinjected with C3 (0.1 mg/ml), GST-RB (2 mg/ ml), MBP-RB/PH (TT) (2 mg/ml), or Rac1^N17 (0.2 mg/ml) along with a marker protein (rabbit IgG, 0.5 mg/ml) into the serum-deprived MDCK cells. After a 30-min incubation, the cells were stimulated with 200 nM TPA for 15 min. (A) The cells into which HA-α-adducin-DD was comicroinjected with GST-RB (panels a and b), MBP-RB/PH (TT) (panels c and d), and Rac1^N17 (panels e and f) are shown. The cells were doubly labeled with TRITC-phalloidin (panels a, c, and e) and FITC anti–rabbit Ig Ab (panels b, d, and f). Arrowheads indicate the microinjected cells. Bar, 25 μm. These results are representative of three independent experiments. (B) The ratios of the membrane ruffling–induced cells to injected cells are indicated. Data are means ± SEM of triplicate determinations.

Effects of α-adducin mutants on the membrane ruffling in the cells introduced with the dominant negative Rho-kinase. MBP, HA-α-adducin (WT), HA-α-adducin-AA, or HA-α-adducin-DD (10 mg/ml) was comicroinjected with C3 (0.1 mg/ml), GST-RB (2 mg/ ml), MBP-RB/PH (TT) (2 mg/ml), or Rac1^N17 (0.2 mg/ml) along with a marker protein (rabbit IgG, 0.5 mg/ml) into the serum-deprived MDCK cells. After a 30-min incubation, the cells were stimulated with 200 nM TPA for 15 min. (A) The cells into which HA-α-adducin-DD was comicroinjected with GST-RB (panels a and b), MBP-RB/PH (TT) (panels c and d), and Rac1^N17 (panels e and f) are shown. The cells were doubly labeled with TRITC-phalloidin (panels a, c, and e) and FITC anti–rabbit Ig Ab (panels b, d, and f). Arrowheads indicate the microinjected cells. Bar, 25 μm. These results are representative of three independent experiments. (B) The ratios of the membrane ruffling–induced cells to injected cells are indicated. Data are means ± SEM of triplicate determinations.