Fig. 4.

Neddylation inhibition enhances the Src-mediated phosphorylation of caveolin-1. a Scratch-based wound healing assays were performed for 24 h in PC3 and U373MG cells which were treated with MLN4924 (0.25 μM and 0.5 μM) or/and 10 μM PP2 (top). The migration areas were calculated using ImageJ at just below. Protein levels in cells lysates were analyzed by Western blotting (middle). The PP2 mediated inhibition of the phosphorylation of Caveolin-1 was quantified based upon the relative level of β-tubulin (bottom). b Transwell migration assays were performed in cells treated with MLN4924 (0.25 μM and 0.5 μM) or/and 10 μM PP2 (left), and migrated cells were counted (right). c Both PC3 and U373MG cells, which had been depleted of NEDD8 using siRNA #1 and si-control, were subjected to wound healing assay for 24 h in the absence or presence 10 μM PP2 (top). The migration areas were calculated using ImageJ at just below. Protein levels in cells lysates were analyzed by Western blotting (middle). The PP2 mediated inhibition of the phosphorylation of Caveolin-1 was quantified based upon the relative level of β-tubulin (bottom). d Transwell migration assays were performed in cells which were prepared as described in C panel (left), and migrated cells were counted (right). Each bar represents the means + standard deviation of results from three independent experiments. * denotes P < 0.05 between the indicated groups. Scale bar = 200 μm