Figure 5

Analysis of the mitogen activated protein kinase pathway (MAPK) in the BxPC-3 and HPAF-II cells. A. Actively growing PC cells were serum starved for 24 h and left untreated or pretreated for 1 h with 5 μg C3 exotransferase, stimulated by the addition of 10% serum and proteins harvested 15 min later. Activated p42/p44 Erk and p38 MAPK were detected by antibodies specific for the phosphorylated forms of each of those proteins. Immunoblots were stripped and reprobed with antibodies specific for total forms of each protein. B. Comparison of active p42/p44 Erk and p38 MAPK in GFP-cav-1 and GFP-vector transient transfectants at 24 h after transfection. C. Results of a colloidal gold migration assay after 30 min pre-treatment of cells with 30 μM of either PD98059 or SB220025. Phagokinetic tracks were imaged at 6 h after stimulation. Results are given for DMSO control (dark grey), PD98059 treated (medium grey) and SB220025 treated (light grey). SB220025 treatment of the HPAF-II cells led to a significant decrease (*p = 0.001) in migration and invasion. Levels of cav-1 protein were increased, while active RhoC was decreased in SB220025 treated HPAF-II cells.