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

Fig. 9.

Fig. 9.

Attenuation of CREB activation leads to down-regulation of P2RX7 agonist-induced c-Fos protein expression. (A) HEK-293 cells transfected with the pIREShyg or pIRES/hP2RX7 expression vectors were stimulated with vehicle (2.5 mM HEPES), 250 μM BzATP, or 1 μg/mL PMA for the indicated times and immunoblotted for total c-Fos protein. Immunoblotting was performed with an anti-Grb2 antibody as a loading control. Analogous results were observed in three separate experiments. (B) HEK-293 cells stably transfected with the pIRES/hP2RX7 expression vector were transiently transfected with MT, S133A (S), or KCREB (K) CREB dn vectors. Twenty-four hours post-transfection, cells were treated with vehicle (2.5 mM HEPES) or 250 μM BzATP for 2 h and immunoblotted for total c-Fos protein expression. Immunoblotting was performed with an anti-Grb2 antibody as a loading control. The results of four independent experiments were collated and are represented as a percent of BzATP-induced c-Fos expression over basal c-Fos expression (mean±sem); *, P < 0.05; **, P < 0.005, as compared with MT-transfected, BzATP-treated c-Fos expression.