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Fig. 1 | BMC Cancer

Fig. 1

From: Inhibition of neddylation facilitates cell migration through enhanced phosphorylation of caveolin-1 in PC3 and U373MG cells

Fig. 1

Migration of some cell-lines was augmented by neddylation block. a PC3 and U373MG cells, which had been transfected with NEDD8 (N8)-targeting (si-NEDD8 #1) or non-targeting (si-Con) siRNAs, were lysed and subjected to Western blotting with antibodies to NEDD8 and β-tubulin (top). The migration potential of the transfected cells were evaluated for 24 h using the scratch-based wound healing analysis. The area of cell migration was calculated using the ImageJ software (middle). The efficiency of the NEDD8 knock down was quantified based upon the relative level of β-tubulin (bottom). b Transwell migration assay was performed to evaluate migration potential of cells. PC3 or U373MG cells, which had been transfected as described in the A panel, were loaded in the upper chamber. After 24 h-incubation, the lower surface of the interface membrane was pictured (top). Cells on the membrane were counted (bottom). Results (means + SDs, n = 3) are presented as relative values vs. the si-Con group. *P < 0.001 vs the si-Con. c Scratch-based wound healing was performed on PC3 and U373MG cells in the presence of MLN4924 (0.25 μM and 0.5 μM) or DMSO as a control. Cell migration was evaluated for 24 h using the scratch-based wound healing analysis (top) and the migration area was calculated using ImageJ (bottom). d Transwell migration assay was performed on PC3 and U373MG cells in the presence of MLN4924 (0.25μM and 0.5μM) or DMSO as a control. After 24 h-incubation, Cells on the lower surface of the membrane were pictured (top) and counted (bottom). All results are presented as the means + standard deviation of three independent experiments, and * denotes P < 0.05 between the indicated groups. Scale bar = 200 μm

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