Effect of Tat-C3 on cell migration. (A) Tat-C3 was preincubated with Raw 264 cells for 30 minutes, and cell migration was assessed in transwell cell culture chambers in the presence or absence of TGF-β1 in the lower chamber. The results are means ± SE of 3 independent experiments, and each value was compared to the corresponding control (Tat-C3, 0 μg/mL) at the same time scale (0, 1, and 24 hours) (*P < .05 and **P < .01). (B) Raw 264 macrophage cells were incubated with TGF-β1 (0-10 μg/mL) for 1 hour, cell lysates (15 μg/mL of total protein) were run on SDS-PAGE, and Western blotting was performed using anti-RhoA antibody. The doublet band consists of unmodified RhoA (lower band) and RhoA modified by Tat-C3 (upper band). (C) Cells were incubated in the upper reservoir with or without a variety of reagents (1 μM cytochalasin-D, 50 μM ML-7, 50 μM Y27632, 10 μg/mL Tat-C3) for 1 hour. The lower chamber was then filled with 5 ng/mL TGF-β1, and migration measured as described in the legend to Figure 1. The values are means ± SE of 3 independent experiments. Each value was compared to the corresponding control at time 0 (***P < .001).