Figure 5. PGE₂ was successfully depleted from RV-induced conditioned medium from HBEC; but it still caused RV-induced β₂ adrenoceptor desensitization and further induced PGE₂ from ASMCs.

HBEC (n = 3) were uninfected (Control) or exposed to: UV inactivated RV (UVi-RV) or replication competent RV (RV) at an MOI = 2 for 24 hours to generate conditioned medium. Conditioned medium was depleted of PGE₂ using affinity chromatography. The original conditioned medium, conditioned medium depleted of PGE₂ and PGE₂ eluted product were collected and the levels of PGE₂ (A) was measured using ELISA. ASMCs (n = 6) were then treated with the original conditioned medium, conditioned medium depleted of PGE₂ or PGE₂ eluted product in BEGM for 3 days. Isoprenaline induced cAMP was measured using a cAMP functional assay (B) and the level of PGE₂ released by ASMCs due to each component was measured using ELISA (C). Data represent mean ± SEM. Statistical differences were detected using a 2-way ANOVA (A&B) and 1-way ANOVA (C) with Bonferroni post test comparisons to the respective control conditioned medium *p<0.05; respective components of the original conditioned medium #p<0.05.