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. 1998 May 12;95(10):5718–5723. doi: 10.1073/pnas.95.10.5718

Figure 2.

Figure 2

Involvement of Ras, MEK1/2, and pp90rsk in MUC2 mucin induction by P. aeruginosa. (A) Effect of coexpressing a dominant-negative mutant form of pp90rsk (pp90rskΔC) on P. aeruginosa-induced MUC2 up-regulation. p-2864luc (15 μg) was cotransfected with a dominant-negative mutant form of pp90rsk into HM3 cells. Forty hours later, HM3 cells were exposed to PAO1 culture supernatant (CS) 6 hr before harvesting. Luciferase activity was measured as described in the text. (B) Effect of PD98059 coexpressing a dominant-negative mutant form and wild-type form of MEK1/2 and a dominant-negative mutant form of SEK [JNKK9(K116R)] on P. aeruginosa-induced MUC2 up-regulation. HM3 cells were cotransfected with p-2864luc (15 μg) and a dominant-negative mutant form (3 μg) or a wild-type form (3 μg) of MEK1/2. Forty hours after transfection, HM3 cells transfected with only p-2864luc were pretreated with PD98059 (37 μM) (Calbiochem) for 30 min. All the cells were then exposed to PAO1 culture supernatant (CS) 6 hr before harvesting. (C) Effect of coexpressing a dominant-negative mutant form of Ras (RasN17) on P. aeruginosa-induced MUC2 up-regulation. HM3 cells were cotransfected with p-2864luc (15 μg) and a dominant-negative mutant form of Ras. The cells were then treated with PAO1 culture supernatant and lysed for luciferase assay. Similar results were observed in another MUC2-expressing epithelial cell line, NCIH292. In the experiments described above, all transfections were carried out in triplicate. Values are the means ± SD; n = 3.