Fig. 6. Effects of various inhibitory agents on mRNA expression in LPS + rDer p2-treated BEAS-2B cells and effects of TLR2 knockdown on the internalization of Der p2 by BEAS-2B cells. (A) Messenger RNA encoding TLR2, IL-6, and IL-8, and β-actin loading control in BEAS-2B cells cultured (6 hours) with rDer p2 in conjunction with LPS. Quantitative data were acquired by densitometric analysis and normalized to the expression of β-actin, which are shown under the bands. (B) Reduced fold induction of IL-6 and IL-8 mRNA induced by rDer p2 in conjunction with LPS. The most effective inhibitors on IL-6/IL-8 mRNA expression were dexamethasone, SP600125 (JNK inhibitor), calcitriol, SB203580 (p38 inhibitor), and BAY 11-7082 (IκB inhibitor). Reduction of fold induction (%) = [1- (LPS + rDer p2 stimulation with inhibitor/without inhibitor)] ×100%. Data are presented as the mean±SEM. (C) TLR2-knockdowned cells were incubated with rDer p2 for 24 hours, followed by rDer p2 protein analysis with immunoblotting. The concentration of Der p2 protein was notably reduced in the cellular lysates of shTLR2-knockdowned and rDer p2-treated cells. A representative image and an actual figure representative of the 3 independent experiments with similar findings are shown. ^*P<0.05; ^**P<0.01.