FIG. 5.

Coactivator LXXLL motif specificity in NF-κB-dependent gene expression. (A) The LXD2 and LXD4 domains of SRC-1/NCoA-1 are required for NF-κB-mediated transcription. Plasmids consisting of a LacZ reporter under the transcriptional control of the E-selectin promoter were injected into the nuclei of Rat-1 cells in the presence of either preimmune IgG or an affinity-purified antibody to SRC-1/NCoA-1. The expression of the reporter plasmid was monitored by X-Gal staining and quantitated based on the percentage of injected cells that stained blue. Rescue experiments were performed by coinjecting the indicated expression plasmids. (B) Coinjection of NCoA-2 expression plasmid does not rescue the inhibitory effect of the anti-SRC-1/NCoA-1 antibody on p65-dependent transcription. (C) Coinjection of NCoA-2 expression plasmid rescues the inhibitory effect of the anti-SRC-1/NCoA-1 antibody on RAR-dependent transcription. In all panels, data are means from three separate experiments; error bars represent standard errors of the means.