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. 2011 Apr 6;31(14):5414–5425. doi: 10.1523/JNEUROSCI.2456-10.2011

Figure 5.

Figure 5.

Requirement for NF-κB-dependent gene expression. A, Fold induction in an NF-κB luciferase reporter assay of 293T cells transfected with the indicated amount (in nanograms) of GFP-tagged wild-type p65 or transcriptionally inactive mutants (p65ΔTAD, p65R33,35A). A reporter containing a mutant NF-κB binding sequence (mκB-lucif) is not activated by transfection with 50 ng of p65, p65ΔTAD, or p65R33,35A. B, Representative confocal projections of dendrites from pyramidal neurons coexpressing mCherry and the indicated GFP or GFP-tagged p65 construct; red and green channels are overlaid to permit visualization of spine enrichment. Scale bar, 10 μm. C, Regions of interest in dendritic spines and shafts were quantitated to calculate normalized dendritic spine enrichment values and percentage enrichment relative to GFP (see Materials and Methods); wild-type p65 and mutant constructs did not significantly differ in spine enrichment. D, Expression of Cre (Cre-IRES-dsRed) in DIV 16 hippocampal pyramidal neurons resulted in a decrease in dendritic spine density that could be rescued by coexpression of GFPp65 but not by the transcriptionally inactive mutants of p65, GFPp65ΔTAD, or GFPp65R33,35A. Dendritic spine density in rescued neurons expressing GFPp65 plus Cre did not significantly differ from control neurons expressing mCherry alone (see text). Data are means ± SEM.