Figure 5.

Salmeterol inhibits MAPK and NF-κB activation. Day 6 DCs were treated with LPS (0.1 µg/ml) and salmeterol (10 µM) for varying amounts of time (0, 15, 30 and 60 min), and total cell lysates were prepared and analyzed for phospho-ERK (1/2) (p-ERK), phospho-JNK (p-JNK) and phospho-p38 (p-p38) expression. Cell lysates were prepared and blotted with the indicated anti-phospho antibodies. Total ERK, JNK and p38 were probed as quantitative controls. Salmeterol inhibits LPS-induced IKKα/β activation and NF-κB nuclear translocation. All proteins were extracted from cells, treated as above, and were western blotted for NF-κB p65. Phosphorylation levels of the proteins were quantified by measuring the band intensities. The results represent three independent experiments. DC, dendritic cell; LPS, lipopolysaccharide; MAPK, mitogen-activated protein kinase; NF-κB, nuclear factor-kappa B.