Fig. 4

OTUD7B inhibits NIK to suppress LCL161-induced NF-κB pathway activation and cell elongation, invasion and migration. a Expression of OTUD7B was determined in several lung cancer cell lines, including H1299, A549, H460, H157, and H1944 cells. b H1299 cells were stably transfected with pcDNA-HA-OTUD7B or empty vector and confirmed by Western blotting. c-h OTUD7B-overexpressing H1299 cells and control cells were treated with 10 μM LCL161 or DMSO for 24 h. c Expression of TRAF3, NIK, p100, p52, and cIAP1 was measured by Western blotting. β-Actin was used as a loading control. d Cell elongation was quantified by calculating the cell elongation index (length/width); the fold increase in the elongation index is shown. e The wound-healing process was monitored by an inverted light microscope. f 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. g NF-kB transcriptional activity was analysed by a luciferase assay, and the fold increase in luciferase activity is shown. h mRNA levels of NF-κB target genes were examined by quantitative RT-PCR. Representative results are shown. Plots are the mean ± SEM of data from three independent experiments. *P < 0.05