Cells overexpressing EphA2 or RacN17, treated with Rho activator or ilomastat acquire an amoeboid- MMP-independent motility style. A) Hs294T, EphA2 or RacN17 transfected cells were serum starved for 48 h, for Ilomastat treatment cells were serum starved for 48 h in the presence of 50 μmol/L Ilomastat, for Rho activation treatment cells were serum starved for 48 h and then stimulated with 1 U/ml Calpeptin for 2 h at 37°C. Then, 6×104 cells were seeded into the upper compartment of Boyden chamber with or without 50 μM Ilomastat. Cells were allowed to migrate through the filter coated with Matrigel toward the lower compartment filled with complete medium. Cell invasion was evaluated after Diff-Quick staining by counting cells in six randomly chosen fields. The results are representative of three experiments with similar results. Student t-test, *p < 0.001 ILO treatment vs untreated, #p < 0.05 ILO treatment vs untreated. B) Analysis of MMP activity. Media from confluent monolayers of cells treated as in A) were collected and analysed by gelatine zymography. The clear bands represent areas of gelatinase activity. The results shown are representative of four experiments.