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. 2011 Aug 31;6(8):e23324. doi: 10.1371/journal.pone.0023324

Figure 4. FANCD2 negatively regulates NF-κB activity and TNF-α transcription.

Figure 4

(A) Overexpression of FANCD2 WT or FANCD2KR-Ub (FANCD2 K561R-Ub fusion), but not the K561R mutant (FANCD2KR), inhibited NF-κB-dependent transcriptional activity. HEK293 cells were transfected with FANCD2 WT, FANCI, FANCG, FANCC, FANCD2 KR, or FANCD2 KR-Ub (each 100 ng) and pNFκB-Luc (100 ng) and pRL-CMV (10 ng). Cells were treated with or without TNF-α(20 ng/ml) for 8 h. Cells were harvested, and dual luciferase assays were performed. Fold activation represents the mean (± s.d) luciferase values of TNF-αstimulated cells normalized with respect to unstimulated cells, from three independent experiments. (B) FA-D2/vec and FA-D2/D2 fibroblast cells were treated with TNF-α (5 ng/ml) for the indicated periods, and real-time PCR analysis of TNF-α mRNA expression in these cells was performed. Gene expression results represent four independent experiments, normalized with respect to unstimulated FA-D2/vec cells, mean (± s.d) from triplicate values are shown.