Figure 3
Bacterial-induced NF-kB DNA binding activity in intestinal epithelial cells. HT-29 cells were untreated, or were exposed to Salmonella typhimurium (1 × 107 colony forming units (CFU)/ml), flagellin (0.5 μg/ml), Clostridium difficile (1 × 107 CFU/ml), or Mycobacterium paratuberculosis (1 × 107 or 1 × 108 CFU/ml) for 1 hr. The DNA binding activity of NF-κB p65 in HT-29 nuclear extracts was determined using an enzyme-linked immunosorbent assay-based transcription factor assay. The positive control Jurkat nuclear extract provided with the kit was used to verify assay specificity in competition assays with wild-type or mutated NF-κB oligonucleotides. The data represent the mean absorbance readings ± standard error of five separate experiments. P < 0.05 relative to untreated cells.