Fig. 4. Cyclooxygenases are involved in IL-4/IL-13 production by IL-33-activated eosinophils.

(A) WT bmEos (5×10⁶ cells/ml) were stimulated with IL-33 (20ng/ml) or nothing (Ctrl). After 2h, mRNA levels of COX-2 were measured by qPCR. (B) WT bmEos were treated with IL-33 (20ng/ml) for 0h, 6h, and 24h respectively, and then protein expression levels of COX-2 were measured by Western blotting. (C and D) WT bmEos (5×10⁶ cells/ml) was treated with IL-33 (20ng/ml) in the presence or absence of indomethacin (50μM) for 24h. Protein levels of IL-4 (C) and IL-13 (D) in the supernatants were measured by ELISA. (E) WT bmEos were treated with sc-560 plus celecoxib (50μM each, COX inhibitors) or vehicle (10μl DMSO) in vitro for 2h, then the cells were washed 3 times and adoptively transferred to ΔdblGATA1 mice followed by APAP injection immediately. Serum levels of ALT were measured at 24h after APAP administration. Two-tailed unpaired Student’s t test was performed in A, and C to E. One-way ANOVA with Tukey post hoc test was performed in B.