FIG. 2.

Coactivator requirements for NF-κB-dependent gene expression. 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 affinity-purified antibodies to the indicated coactivators. The expression of the reporter plasmid was monitored by staining with 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside (X-Gal) and quantitated based on the percentage of injected cells that stained blue. (A) Effects of nuclear microinjection of anti-CBP, SRC-1/NCoA-1, and pCAF antibodies on p65-induced E-selectin-lacZ reporter gene expression. Photomicrographs of rhodamine-stained injected cells (top panel) and the corresponding phase-contrast pictures (lower panel) display typical results. (B) Coactivator requirements of NF-κB-dependent gene expression. Results were repeated in three separate experiments with more than 200 cells injected for each data point; data are expressed as means, and error bars represent standard errors of the means. (C) Effect of nuclear microinjection of anti-CBP, SRC-1/NCoA-1, and p/CAF antibodies on Sp1-LacZ and CMV-LacZ reporter constructs.