Figure 6

eIF6 is implicated in the control of cell motility/invasiveness in WM793 melanoma cancer cells inducing an increased expression of cdc42 protein. The results of eIF6 over-expression on the migratory activity of the cells were generalizable to other cell line models. A) eIF6 and cdc42 expression in WM793 primary melanoma cell lines transiently transfected either with the pcDNA3.1-eIF6 or control plasmid was analyzed by western blotting. B) qPCR of cdc42 mRNA was performed analysing 2 μg of total RNA reverse-transcribed into cDNA and comparing its expression between WM793 primary melanoma cell lines over-expressing eIF6 with respect the control. The bar graphs represent the relative fold changes of cdc42 mRNA presented as mean ± S.D. and relative to that of GAPDH. The results are the average of three independent experiments C) Migration assay: WM793/eIF6 and control cells were seeded in the upper side of migration chambers. The cells migrated to the lower chamber after 48 h of incubation were stained with crystal violet dye. D) Invasivity assay: cells were seeded in the upper side of invasion chambers. After 48 h cells migrated in the lower chamber were stained with crystal violet dye. The total stained area in the lower chambers was estimated using the Image-J software. The cell images in C and D are representative of three independent experiments. The histograms in B, C and D represent the average of three independent experiments. The P-values were calculated with the t-test using the symbols (**) and (*) corresponding to < 0.05 and < 0.01, respectively.