Fig. 5. p65-NFB nuclear export is required for cytoplasmic retention of N-CoR. (A) Immunofluorescence staining of 293T cells transfected with N-CoR-flag and p65TA in the absence (upper panel) or presence (lower panel) of LMB (10 ng/ml). (B) Immunofluorescence staining of 293T cells transfected with N-CoR-flag and p65NES. N-CoR was detected with an -flag antibody and a FITC-conjugated secondary antibody (left panels), -p65 staining is represented in the middle panels corresponding to Cy3 signal, and nuclei were visualized by DAPI staining (right panels). (C) Hes1-luc transcriptional activity of NIH-3T3 cells cotransfected with N1-IC, N-CoR or/and p65TA plasmids as indicated. Luciferase activity is represented as the fold induction relative to the basal level measured in cells transfected with empty vector. The average and standard deviation of duplicates from a representative experiment are presented. Lower panels show p65 protein levels. Protein loading for each lane was normalized according to ß-galactosidase expression.

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