FIG. 5.

CREB binds to NE and activates transcription of a downstream reporter gene. (A) Sequence alignment of the CRE and NE. (B) CREB recognizes the NE in EMSAs. EMSAs to detect supershifts were performed in the presence of anti-CREB and anti-P-CREB antibodies. The identities of the probes and the amounts of antibody and incubation periods are labeled at the tops of the lanes. (C) P-CREB transactivated both CRE-luc and NE-PCE-luc reporter genes in transiently transfected NIH 3T3 cells. CRE-luc and NE-PCE-luc reporter genes were cotransfected with CREB, CREB plus PKA, or K-CREB plus PKA into 3T3 cells. Sample values are means ± standard errors of results for eight samples. All results were normalized to the relative luciferase activities in cells transfected with the CRE-luc reporter gene plus CREB, which was assigned a value of 1.0. ***, P < 0.001; **, P < 0.01 (the value was significantly different from that for the respective control [CREB only]).