Fig. 2.—
Effect of kinase inhibitors on MUC5AC protein production induced by Pseudomonas aeruginosa culture supernatant (PA Sup) and by epidermal growth factor (EGF) in NCI–H292 cells. Confluent NCI–H292 cells were stimulated with M9 medium (1:8 dilution), with Roswell Park Memorial Institute (RPMI) medium alone (control), with EGF (positive control; 5 ng·mL−1), or with PA Sup (1:8 dilution) in serum-containing (fig. 2a⇓) or in serum-free (fig. 2b⇓) RPMI medium for 24 h. To examine the effects of kinase inhibitors, NCI–H292 cells were pretreated with BIBX1522 (BIBX, 10 µM), tyrphostin AG1478 (AG1478, 10 µM), PD98059 (30 µM), tyrphostin AG1295 (AG1295, 10 µM), or tyrphostin A1 (A1, 10 µM). Upon completion of the treatment, the cell lysate and the cell culture supernatant were collected, and total amount of MUC5AC protein was measured by enzyme-linked immunosorbent assay. ƒ: M9; ##: EGF; ¶¶: PA sup. Results are reported as a) % above serum-containing control value or b) % above serum-free control value (mean±sem; n=5, for each condition). **: p<0.01, ***: p<0.001, significantly different from serum-containing control; #: p<0.05 significantly different from serum-free control;¶: p<0.01 significantly different from corresponding stimulated state; +: p<0.001, significantly different from corresponding stimulated state; §: p<0.05 significantly different from PA Sup alone.