Fig. 2.

The induction of NOXO1 expression in gastric cancer cells by TNF-α/NF-κB pathway. a The relative mRNA levels of NOXO1 in the TNF-α-stimulated gastric cancer cells SNU601, SNU719, MKN45, and KATOIII are shown (mean ± s.d.). *p < 0.05; **p < 0.01. b Immunoblotting analyses for NOXO1 (left) and phosphorylated p65 (right) in the SNU601 cells after TNF-α stimulation at the indicated time. The relative band intensities to 0 h of TNF-α stimulation are indicated at the top of the panels, along with graphs after normalization with GAPDH (left) or p65 (right). c The relative mRNA levels of RELA (left) and NOXO1 (right) normalized with the levels of GAPDH in the TNF-α-stimulated or unstimulated SNU719 cells are shown (mean ± s.d.). *p < 0.05; N.S. not significant. d A schematic view of the NOXO1 promoter regions (from −2000 bp to the transcription start site [TSS]) showing two putative NF-κB binding sites, indicated by boxes. Mut1 and mut2 vectors contain mutations at either of the two NF-κB binding sites. These fragments were used for a luciferase reporter assay. e The relative luciferase activities in SNU601 cells after TNF-α stimulation at 0, 3, and 6 h are shown (mean ± s.d.). ***p < 0.001. f The relative percent inputs of ChIP-based real-time PCR results for NOXO1 promoter in SNU601 cells after TNF-α stimulation at 0 and 3 h are shown (mean ± s.d.). ***p < 0.001; N.S. not significant