Figure 5.

Inhibition of P. aeruginosa-induced MUC 2 up-regulation by tyrosine kinase inhibitors. (A) HM3 cells were or were not pretreated with genistein (Genist) (100 μg/ml) (Sigma) for 2 h and then were exposed to P. aeruginosa PAO1 culture supernatants (PA) or vehicle (Con) for 6 h before RNA extraction and RPA analysis. The results are typical of four separate experiments. (B) HM3 cells were transfected with p-2864luc. After 40 h, the cells were pretreated with genistein (100 μg/ml) for 2 h and then were exposed to P. aeruginosa culture supernatants (PAO1) for 6 h before harvesting for luciferase analysis. (C) Forty hours after being transfected with p-2864luc, HM3 cells were pretreated with tyrphostin AG 126 (25 μM) (Calbiochem) for 3 h and then were exposed to PAO1 culture supernatant (CS) or LPS from PA10 (serotype 10) (5 μg/ml) for 6 h before harvesting. Luciferase activity was measured as described above. All transfections in B and C were carried out in triplicate. Values represent means ± SD (n = 4). Luciferase activity was normalized with respect to β-galactosidase.