Fig. 3

NIK is required for LCL161-induced NF-κB pathway activation and cell elongation, invasion and migration. H1299 cells were transfected with shRNA against NIK or the control vector for 48 h and then treated with 10 μM LCL161 or DMSO for 24 h. a Expression of NIK, p100, p52, and cIAP-1 was analysed by Western blotting. β-Actin was used as a loading control. b Cell morphology was analysed by phase-contrast microscopy. Magnification, × 100. Cell elongation was quantified by measuring cell length and width and by calculating the cell elongation index (length/width), and the fold increase in the elongation index is shown. c The wound-healing process was monitored by an inverted light microscope. d Cells were plated in a collagen-coated transwell migration chamber. After treatment, the migrated cells were fixed and stained with crystal violet, and optical density was measured. e The mRNA levels of NF-κB target genes were evaluated by quantitative RT-PCR. Representative results are shown, and plots represent the mean ± SEM of data from three independent experiments. *P < 0.05