Figure 7.

Apremilast suppressed the inflammatory responses in macrophages through PKA–CREB signalling. (a) TNF‐α and IL‐10 production in LPS‐stimulated BMDMs and RAW264.7 cells. (b) The phosphorylation of CREB and ATF‐1, following apremilast treatment, determined by western blot. (c) Immunofluorescent staining for phospho‐CREB upon apremilast treatment. BMDMs and RAW264.7 cells were treated with 10‐μM apremilast, 10‐μM H89 (PKA inhibitor), and 10‐μM Forskolin, cAMP level (d), phosphorylation of CREB and ATF‐1 (upper e, upper f, and g) were measured and TNF‐α production were determined upon LPS stimulation (h). BMDMs and RAW264.7 cells were transfected with PKA siRNA and NC SiRNA; then cells were treated with apremilast, H89, and forskolin. The phosphorylation of CREB and ATF‐1 (lower part of e; lower part of f) and TNF‐α (i) were measured. The summary data are shown as means ± SEM of three independent experiments