Fig. 6. LPA-induced changes in actin cytoskeleton and focal contact organization. PANC-1 cells were starved overnight in DMEM + 0.1% BSA, wounded and treated for 3 hours with DMEM supplemented with 0.1% BSA (a,b), 10 µM LPA (c,d), 10 µM LPA, 25 ng/ml PTX (e,f), or 10 µM LPA, 25 µM PD98059 (g,h). PTX and PD98059 were added 16 hours and 15 minutes, respectively, prior to wounding. Paraformaldehyde-fixed cells were costained with TRITC-conjugated phalloidin to visualize filamentous actin and with anti-vinculin and Alexa488-conjugated antibodies to visualize vinculin. Microscopy was performed with an inverse fluorescence microscope. Arrows in panel c point to actin-rich lamellipodia, arrows in panel d point to focal contacts. Images in g' and h' represent enlargements of the indicated sections in g and h, respectively. The illustration in i shows the merged images of g' and h'. The yellow color indicates overlapping of red and green structures. Bars, 25 µm.