Fig. 5

Overexpression of 14-3-3 eta inhibits Ras-induced focus formation in NIH3T3 cells. a H-ras and Flag-14-3-3 eta expression plasmids were transfected into NIH3T3 cells as described in the Methods section, replated 24 h after transfection and maintained for an additional 21 days. Subsequently, the cells were stained with crystal violet and the number of foci quantified using an Oxford Optronix colony counter, UK. The experiment was conducted in octuplicate. Representative plates from each of the four experimental groups are depicted. b In a parallel control experiment NIH3T3 cells were transfected with H-ras and H-ras plus empty vector and treated as in a. The experiment was conducted in octuplicate. c The number of foci in panels a and b were quantified and plotted. Horizontal bars depict the average number of foci per dish. (*: p < 0.05, Student’s t-Test). d A western blot was used to verify the expression of H-ras and N-terminally-tagged Flag-14-3-3 eta proteins in NIH3T3 transformants. e A western blot was used to verify expression of H-ras in both Ras and Ras plus Vector group transformants shown in panel b