Figure 2
BaP1 activates COX-1 and -2 pathways to release PGE2 and up-regulates COX-2 expression. (A) Isolated FLSs were pretreated with the selective inhibitors of COX-1 and -2, valeryl salicylate and NS398, respectively, for 1 h and incubated with BaP1 (12.5 µg/mL) or RPMI (control) for 3 h. Supernatants were collected, and PGE2 was quantified by ELISA. (B–E) FLSs were incubated with BaP1 (12.5 µg/mL) or RPMI (control) for 30 min, 1, 3 and 6 h. (B) Representative Western blotting of COX-1 and β-actin (loading control) showing immunoreactive bands. (C) Representative Western blotting of COX-2 and β-actin (loading control) showing immunoreactive bands. Densitometric analysis of immunoreactive (D) COX-1 and (E) COX-2 bands. (F) COX-2 relative gene expression determined by quantitative PCR in cells incubated with BaP1 (12.5 µg/mL) or RPMI (control) for 15 min, 3 and 6 h. Data are mean ± SEM of 3 samples. *p < 0.05 relative to control; #p < 0.05 relative to BaP1 (with vehicle); &p < 0.05 relative to BaP1 (6 and 12 h) (one-way ANOVA, Bonferroni post test in A and two-way ANOVA, Bonferroni post test in D, E and F). Blots were cropped from the full-length blots, which are demonstrated as Supplementary Figures 3(A,B) and 3(A–D) and cropped blots are indicated by arrows.
