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. 2016 Jan 27;6:19922. doi: 10.1038/srep19922

Figure 2. Promotion of Ad early gene expression and replication by TNF-α stimulation.

Figure 2

(A) HeLa cells were transfected with pNF-κB-Luc, followed by treatment with recombinant hTNF-α at 100 ng/ml. After 24-h incubation, luciferase activity was determined. The data show FLuc activity normalized by RLuc activity. (B,C) HeLa cells were pre-treated with hTNF-α at 100 ng/ml for 5 h, followed by infection with WT-Ad at 100 VP/cell. After 12-h incubation, the E1A, E2, E3, and E4 mRNA levels in the cells were determined by quantitative RT-PCR (B). After 24-h incubation, the E1A protein levels in the cells were determined by western blotting analysis (C). The intensity of E1A expression was quantified using Image J software. (D) HeLa cells were pre-treated with hTNF-α at the indicated concentration for 5 h, followed by infection with WT-Ad at 100 VP/cell. After 24-h incubation, Ad genome copy numbers in the cells were determined by quantitative PCR. (E) HeLa cells were pre-treated with hTNF-α at 100 ng/ml for 5 h, followed by transduction with Adv-CMVLuc at 100 VP/cell. After 12-h incubation, Ad gene mRNA levels in the cells were similarly determined. These data are expressed as the means ± S.D. (n = 3–4). *p < 0.05, **p < 0.01, ***p < 0.001.