Figure 3.

CLAN formation in DEX-treated HTM cells is dependent on Rac1/Trio signaling. HTM cells were treated with 0.1% ethanol or 500 nM DEX for 4 days and then plated and spread on fibronectin for 1.5 to 2 hours in the absence (A) or presence (B) of mAb AP-5 ± the PI-3 kinase inhibitor LY294002 or the Rac1/Trio inhibitor NSC23766. Cells were fixed and labeled with phalloidin, and the percentage of CPC was determined. Results are represented as the mean ± SD. (A) The increase in the DEX only-treated cells relative to control cells and the decrease due to treatment with the Rac1 inhibitor relative to its control were statistically significant at P < 0.01. There were no statistically significant differences between the other treatment groups. n ranged from 1649 to 1863 cells. (B) The decrease in AP-5-induced CLAN formation after treatment with the NSC23766 inhibitor was statistically significant at P < 0.01 in both DEX-treated and ethanol-treated cells. In the presence of DEX, the decrease in AP-5-induced CLAN formation with the LY294002 inhibitor was statistically significant at P < 0.01 relative to the DMSO-treated control. In the presence of ethanol only, the decrease in AP-5–induced CLAN formation after treatment with the LY294002 inhibitor was statistically significant at P < 0.05 relative to the untreated control. There was no difference between cells treated with ethanol and AP-5 relative to ethanol, AP-5, and DMSO or between cells treated with ethanol, AP-5, and DMSO relative to those treated with ethanol, AP-5, and LY294002. n ranged from 1441 to 1576 cells.