Figure 4.

Effect of MK2206 on EEC4 and AN3CA cells in vitro. (A) Cells from EEC4 and AN3CA were cultured in vitro and treated with increasing concentrations of MK2206 (0, 0.01uM, 0.1uM, 1uM, 10uM). Levels of p(Ser473)-AKT, p(Thr308)-AKT and pan AKT were measured by western blot. (B) EEC4 and AN3CA were plated on matrigel coated membranes and treated with 1uM MK2206 for 36h and 72h. Cells on top were scraped off and cells that invaded through to the underside of the membrane was visualized with Diff-Quik stain. Invading cells were visualized under light microscopy and representative pictures are shown of cells treated with 1uM MK2206 for 72h (magnification 200X). The number of invading cells was counted in 4 different fields using ImageJ and the average was taken. Data was log transformed for statistical analysis. Data are presented as percent changes of MK2206 treated compared with vehicle treated cells. (* denotes statistical significance, p < 0.05).