Figure 5.

TNF-α induces MACC1 via c-Jun/AP1. (A) HCT116 cells were transiently transfected with the MACC1 promoter reporter plasmids with either mutated AP-1 or Sp1 transcription factor binding sites along with a renilla luciferase control plasmid for 24 h. Cells were then treated with increasing concentrations of TNF-α. After 24 h of TNF-α treatment luciferase activity was measured and normalized to renilla luciferase activity. (B,C) HCT116 cells were transfected with the target-specific predesigned c-Jun siRNA or scrambled control siRNA for 24 h. Cells were then stimulated with increasing concentrations of TNF-α for another 24 h. Cells without TNF-α treatment served as controls. Cells were analyzed to assess the c-Jun and MACC1 mRNA and protein expression levels using qRT-PCR and Western blotting, respectively. The data is presented as mean ± SEM with the statistical significance levels: **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001.