FIGURE 5.

Dominant negative ARF expression, β-arrestin depletion, and truncated mutant expression inhibit Ang II-dependent cell migration. A–C, HA-AT₁R-expressing HEK 293 cells were transfected with ARF6 T27N for 24 h (A), with control (ctl) or a mixture of β-arrestin1 and -2 siRNA for 48 h (B), or with an empty vector, full-length β-arrestin2, or a truncated mutant (β-arrestin2-(1–320)) for 24 h (C). Cells were trypsinized and reseeded into Boyden chambers where they were left for 1 h. One set of cells was stimulated with Ang II (100 nm), and the other was left untreated. Migration to the lower chamber was evaluated after 4 h for all conditions. D, HA-AT₁R-expressing HEK 293 cells were trypsinized and reseeded in Boyden chambers where they were pretreated with DMSO or PitStop2 (10 μm) for 30 min. One set of cells was stimulated with Ang II (100 nm), and the other was left untreated. Migration to the lower chamber was evaluated after 4 h for all conditions. For all experiments, quantifications are the mean ± S.E. of three independent experiments. ***, p < 0.001; **, p < 0.01; *, p < 0.05. ns, nonstimulated.