We previously reported that p38 MAPK signaling is required for osteoclast differentiation but not osteoclast function. Here we further investigated the role of p38 MAPK in the function and differentiation of mouse bone marrow macrophages (BMM phi), common precursors of osteoclasts and dendritic cells. Lipopolysaccharide (LPS) activated the p38 MAPK signaling pathway in BMM phi by sequential phosphorylation of MAPK kinase 3/6, p38 MAPK, and activating transcription factor-2. Treatment of BMM phi with SB203580, a p38 MAPK inhibitor, suppressed LPS-induced phosphorylation of activating transcription factor-2. LPS stimulated production of IL-1 beta, TNF alpha, and IL-6 in BMM phi, and SB203580 failed to inhibit the LPS-induced cytokine production. BMM phi incorporated latex beads via phagocytosis, and SB203580 had no effect on this phagocytosis. BMM phi differentiated into dendritic cells when treated with granulocyte macrophage colony-stimulating factor together with CD40 ligand, TNF alpha, or LPS, and SB203580 failed to inhibit this differentiation. Thus, p38 MAPK-mediated signals are not involved in either BMM phi function or BMM phi differentiation into dendritic cells. The differentiation of BMM phi into osteoclasts in response to receptor activator of nuclear factor-kappa B ligand or TNF alpha was strongly inhibited by SB203580. These findings emphasize the crucial roles of p38 MAPK-mediated signaling in osteoclast differentiation.