Profiling protein phosphorylation in RAP-treated BMDMs. (A) BMDMs from WT C57BL/6J mice were treated with RAP (150 nM) or vehicle for 1 h. Protein phosphorylation was analyzed by array. (B) Changes in protein phosphorylation associated with RAP treatment in comparison with cells treated with vehicle. Each of the analyzed phoshoproteins is demarcated with a box in panel A. (C) BMDMs were stimulated for 1 h with increasing concentrations of RAP (10-300 nM) and with RAP that had been boiled for 5 min (150 nM). Cell extracts were subjected to immunoblot analysis to detect phospho-ERK1/2 and total ERK1/2. (D) BMDMs were treated with LPS (0.1 µg/mL), RAP (150 nM), or vehicle for 1 h. Cell extracts were subjected to immunoblot analysis to detect phospho-p38 MAP kinase, total p-38 MAP kinase, phospho-JNK, and total JNK.