Fig. 3

Overexpression of eIF-5A2 induces cell proliferation and migration, and enhances translation. A Protein extracts from empty vector (H1395-EV) or Flag-eIF-5A2 (H1395-eIF-5A2) stably-transfected H1395 cells were analysed by western blot with eIF-5A2 and Flag antibodies. HSP90 was used as a loading control protein expression. A representative image of the three idependent experiments is shown. B Cell proliferation assay in H1395-EV and H1395-eIF-5A2 cells. MTS assays were performed 24 h, 48 h and 72 h after seeding the cells. Experimental means (n = 6 with experimental triplicates) were compared by two-way ANOVA with Tukey's test for multiple sample comparison (**p < 0.01). C Cell migration assay in H1395-EV and H1395-eIF-5A2 cells. Wound was performed in confluent monolayer cell culture, and wound closure was monitored by phase-contrast microscopy. The means of the experiments (n = 3 with experimental triplicates) were compared by two-way ANOVA analysis with Tukey's test for multiple comparison of samples (*p < 0.05) (right panel). Representative images of phase contrast microscopy are shown in the left panel. Scale bar 500 µm. D Polysomal profile of H1395-EV and H1395-eIF-5A2 cells. Global RNA polysome profiles generated by measurement of the OD at 254 nm in the density gradient fractionation system are shown. From left to right, each peak represents the 40S subunit, the 60S subunit, monosomes (80S), disomes, and polysomes. A representative graph of the experiments performed is shown (n = 3). E Protein extracts from the fractions obtained in D were analysed by western blot with eIF-5A2, RPL4 and rpS6 antibodies. A representative image of the experiments performed is shown (n = 2)