Fig. 5: NF-κB pathway contributes to the upregulation of genes related to angiogenesis and cell migration.

Ranking of the top ten upregulated transcription factors presumed to be involved in C4OE (a) and C4KO (b) by TRRUST database analysis of the CDS expression data obtained from RNA sequencing of NCI-H292 cells. The horizontal axis shows accumulative hypergeometric p values. c Comparison of relative luciferase activity between mock and C4KO NCI-H292 cells. d Immunoblotting analysis of phospho-p65, p65, and CASP4 in mock and C4KO cells. e Comparison of relative luciferase activity between mock and C4KO cells with rescued CASP4 gene (C4KO + C4) (n = 3). f Immunoblotting analysis of phospho-p65 and CASP4 protein in mock, C4KO, and C4KO + C4 cells. g mRNA levels of CASP4, PTGS2, SERPINE1, and EPHA2 in mock and C4OE cells with or without treatment with the NF-κB inhibitor BAY11-7082. h Protein levels of CASP4 in mock and C4OE cells with or without treatment with BAY11-7082. i Representative images of the wound healing assay with or without treatment with BAY11-7082 using mock or C4OE NCI-H292 cells at 0 and 24 h (scale bar, 500 μm). j The ratios of wound width after 24 h to initial wound width in (i) were graphed as percent wound closure (n = 3). In each experiment, treatment with BAY11-7082 (2.5 μM) lasted 8 h. n.s.: not significant. Data are expressed as mean ± standard deviation (n = 3). *P < 0.05, **P < 0.01 compared with mock. Densitometry values are shown above each blot.