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. 2008 Jul 14;84(4):1159–1171. doi: 10.1189/jlb.0907612

Fig. 4.

Fig. 4.

Effect of an ERK1/2 kinase inhibitor on P2RX7-induced CREB phosphorylation in murine macrophages. (A) RAW 264.7 macrophages were pretreated for 15 min with vehicle (DMSO), the MEK1/2 inhibitor UO126 (10 μM), or the MEK1/2 inactive inhibitor analog UO124 (10 μM). The cells were subsequently treated with vehicle (2.5 mM HEPES) or 250 μM BzATP for 15 min. A representative immunoblot displaying pCREB/pATF1 is shown. Immunoblotting was performed with anti-pan-ERK1/2 antibody as a loading control. The results of five independent experiments were collated and are represented as normalized CREB phosphorylation (mean±sem); *, P < 0.00001. (B) RAW 264.7 macrophages were pretreated for 15 min with vehicle (DMSO) or UO126 at concentrations from 1 μM to 30 μM. The cells were subsequently treated with vehicle (2.5 mM HEPES) or 250 μM BzATP for 15 min. A representative immunoblot displaying pCREB/pATF1 is shown (n=3). (C) RAW 264.7 macrophages were pretreated for 15 min with vehicle (DMSO) or 10 μM UO126. The cells were subsequently treated with vehicle (2.5 mM HEPES) or 3 mM ATP for 15 min. The results of three independent experiments were combined and are represented as a percent of ATP-induced CREB phosphorylation (mean±sem); **, P < 0.05.