Figure 4

Nuclear factor- κB (NF- κB) p50 and p65 bind more efficiently to the 5 T and 3T5T long terminal repeats (LTRs). Nuclear extract was isolated from normal and tumor necrosis factor-α (TNF-α)-treated TF-1 cells. (A) An LAI long probe covering CCAAT/enhancer binding protein (C/EBP) site I, NF-κB site II, NF-κB site I, and Sp site III with the wild-type (WT), 3 T, 5 T, or 3T5T single-nucleotide polymorphisms (SNPs) was used to determine differences in complex formation at each LTR. TF-1 nuclear extract was incubated with probe in the absence or presence of (B) NF-κB p50 or (C) p65 gel shift antibodies for 30 minutes. (B) Normal and activated extracts incubated with the 5 T and 3T5T long probe showed greater complex formation and abrogation and shift of complex formation when incubated with NF-κB p50 antibody (black arrows). (C) Normal and activated extracts incubated with the 5 T and 3T5T long probe showed greater complex formation and abrogation of complex formation when incubated with NF-κB p65 antibody (black arrow).