Figure 4.

Acanthamoeba plasminogen activator–upregulated IL-8 is not diminished by PAR1 antagonist. HCE cells were incubated with aPA (100 μg/mL) and PAR1 agonists (thrombin, 10 μM; TRAP-6, 10 μM) for 48 hours. Inhibition of PAR1 involved preincubating the HCE cells for 1 hour with the PAR1 antagonist (SCH 79797, 60 μM) and then incubating with or without aPA or PAR1 agonists for 48 hours. (A) Total RNA was isolated and assessed using qRT-PCR for mRNA expression of IL-8. Relative fold change of mRNA expression was quantified by 2^−ΔΔCt method.³⁵ (B) Supernatants were collected from harvested cells and subjected to IL-8 ELISA. The data are mean ± SEM of three independent experiments (*P < 0.05). P values were obtained by unpaired Student's t-test.