A) HEK293 cells were transfected with the indicated amounts of pCMV-Myc vector control, P2X7 WT/pCMV-Myc or P2X7 N187A/pCMV-Myc. After 24 h, the cells were treated with 250 μM BzATP for 5 min, and cell lysates were prepared and immunoblotted with anti-Myc, anti-pERK1/2 and anti-ERK1/2 antibodies. These data are representative of two independent experiments. B) COS7 were transfected with pCMV-Myc, P2X7 WT/pCMV-Myc, P2X7 N187A/pCMV-Myc or P2X7 N202A/pCMV-Myc. After 24 h, a YO-PRO-1 dye uptake was performed. Similar data were obtained in two separate experiments. C) HEK293 cells were transfected with pCMV-Myc vector control, P2X7 WT/pCMV-Myc or P2X7 N187H/pCMV-Myc. After 24 h, the cells were treated with 250 μM BzATP for 5 min, and cell lysates were prepared and immunoblotted with anti-Myc, anti-pERK1/2 and anti-ERK1/2 antibodies. These data are representative of three independent experiments. D) HEK293 cells were transfected with pCMV-Myc vector control, P2X7 WT/pCMV-Myc or P2X7 N187Q/pCMV-Myc. After 24 h, the cells were treated with 250 μM BzATP for 5 min, and cell lysates were prepared and immunoblotted with anti-Myc, anti-pERK1/2 and anti-ERK1/2 antibodies. These data are representative of three independent experiments.