Figure 2.

Injury increases the levels of CRE-binding proteins. (A) As reported previously, pedal ganglia extracts give rise to three specifically retarded bands (arrowheads) when examined by using EMSAs (40). The specificity of the CRE sequence-containing probe for CRE-binding proteins was determined by competition EMSAs. The inclusion of a different, unlabeled CRE sequence-containing oligonucleotide effectively abolished the binding to the CRE probe at both ×25 and ×100 molar excess. In contrast, an oligonucleotide containing a mutated CRE sequence (CREmut) but identical flanking sequences to the probe did not compete for binding. (B) Picture of a representative EMSA obtained using protein extracts from control and 3-hr postinjury pedal ganglia samples. As described in Experimental Procedures, samples prepared from the uninjured symmetrical pedal ganglia were used as controls for each injured sample. (C) Summary figure showing that at 3 hr postinjury, the optical density of retarded band 1 is increased significantly as compared with the controls. The data are presented as the mean ± SEM. ∗, P < 0.05. (D) Picture of a representative EMSA obtained by using protein extracts from control and 24-hr postinjury pedal ganglia samples. (E) Summary figure indicating that at 24 hr postinjury, the optical density for the retarded band 2 is increased significantly as a result of injury. The data are presented as the mean ± SEM. ∗, P < 0.05.