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

Fig. 2.

Fig. 2.

Dependence of nucleotide-induced pCREB on P2RX7 function/expression. (A) Wild-type (WT) and serine to phenylalanine mutation (SF) mutant RAW 264.7 macrophages were treated with vehicle (2.5 mM HEPES), 10 μg/mL anisomycin, 1 μg/mL LPS, 250 μM BzATP, or the indicated concentrations of ATP for the time-points shown. A representative immunoblot displaying pCREB/pATF1 is shown (n≥3). Immunoblotting was performed with anti-pan-ERK1/2 antibody as a loading control. (B) Untransfected HEK-293 cells and HEK-293 cells transfected with the pIREShyg (Hyg) or pIRES/hP2RX7 (P2RX7) expression vectors were stimulated with vehicle (2.5 mM HEPES), 10 μg/mL anisomycin, 250 μM BzATP, or 100 μM forskolin for the indicated times and immunoblotted for pCREB/pATF1. Immunoblotting was performed with anti-pan-ERK1/2 antibody as a loading control. Results of the three independent experiments were collated, and BzATP-induced pCREB was graphed ± sem.