Fig. 3. αDgk dominant-negative mutant and R59949 inhibit HGF-induced cell motility of PAE cells. (A) Quiescent PAE cells infected with either PINCOS (open boxes), PINCOS/myc-αDgk (hatched boxes) or PINCOS/myc-αDgkK^– (dark boxes) retrovirus were induced to migrate in a Boyden chamber assay with 250 U/ml HGF or 10% FCS in the presence or absence of 1 µM R59949. The graph shows a representative experiment of three. Each value is the mean ± SE of triplicates. The differences between HGF-stimulated PINCOS versus HGF-stimulated PINCOS/myc-αDgkK^–, HGF-stimulated PINCOS in the presence versus absence of 1 µM R59949 and HGF-stimulated R59949-treated PINCOS versus PINCOS/myc-αDgk are statistically significant (t-test, p <1%). (B) Quiescent PAE cells infected with either PINCOS, PINCOS/myc-αDgk or PINCOS/myc-αDgkK^– were solubilized in Laemmli buffer, 50 µg of total proteins were separated by 10% SDS–PAGE and analyzed by western blotting with αDgk antibodies.