Fig. 5

Knockdown of SCEL expression significantly decreases TNF-α-induced NF-κB activation and the expression of NF-κB-driven anti-apoptotic genes. A Western blotting analysis of p65 activation upon TNF-α stimulation in multiple SCEL stable knockdown LC clones and the control LC cells. B Analysis of NF-κB reporter assay using the SCEL stable knockdown LC cells and the control LC cells. *P < 0.001. C Analysis of TNF-α-dependent p65 activation kinetics in SCEL-downregulated LC cell and the control LC cells. *P < 0.01. D The representative IF images of TNF-α-induced p65 phosphorylation and nuclear translocation in the SCEL-downregulated LC cells and the control LC cells. Scale bar, 20 μm. E Quantitative result of phosphorylated p65 nuclear translocation. *P < 0.01. F Analysis of nuclear translocation of phosphorylated p65 in the control cells and the SCEL-downregulated cells upon TNF-α stimulation using subcellular fractionation. G qRT-PCR analysis of anti-apoptotic gene expression in response to TNF-α-stimulation in the control LC and SCEL-downregulated LC cells. H qRT-PCR analysis of proliferation-related gene expression in response to TNF-α-stimulation in the control LC and SCEL-downregulated LC cells. *P < 0.01. Each experiment was performed in triplicates and was repeated at least 3 times