FIGURE 2.

Cytokine (IL6 and IL8) levels in cell culture supernatants. HT-29 cells were grown in 6-well plates to 50% confluence and serum-starved for 24 h. Cells were then treated for 48 h with trefoil (10 μg/ml). In some experiments, HT-29 cells were preincubated for 24 h with LPS (1 μg/ml) before treating with trefoil for 48 h. Afterwards, culture supernatants of HT-29 were collected and centrifuged at 1000g for 15 min at 4°C, and proteins were precipitated by TCA. Concentrations of cytokines were determined by ELISA using anti-IL6 or anti-IL8 as primary antibody as indicated in “Materials and Methods”. The figure shows the measured concentrations of IL6 (A) and IL8 (B) expressed as μg/ml. Each assay was carried out in three independent experiments, and the results are reported as mean ± S.D.