Effect of MAGI proteins knockdown on CRF-mediated ERK1/2 signaling. (A) Representative immunoblot showing ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min in non-transfected (NT) HEK293 cells, and HEK293 cells transfected with HA-CRFR1 along with either SCR siRNA or MAGI-1 siRNA. Also shown are corresponding immunoblots for total ERK1/2, MAGI-1 and HA-CRFR1 protein expression. (B) Densitometric analysis of ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min. The data represent the mean ± SEM of four independent experiments. (C) Representative immunoblot showing ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min in non-transfected (NT) HEK293 cells, and HEK293 cells transfected with HA-CRFR1 along with either SCR siRNA or MAGI-2 siRNA. Also shown are corresponding immunoblots for total ERK1/2 and HA-CRFR1 protein expression. (D) Densitometric analysis of ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min. (E) Representative immunoblot showing ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min in non-transfected (NT) HEK293 cells, and HEK293 cells transfected with HA-CRFR1 along with either SCR siRNA or MAGI-3 siRNA. Also shown are corresponding immunoblots for total ERK1/2, MAGI-3 and HA-CRFR1 protein expression. (F) Densitometric analysis of ERK1/2 phosphorylation in response to 500 nM CRF treatment for 0, 5, and 15 min. The data represent the mean ± SEM of four independent experiments. *P < 0.05 versus SCR siRNA control.