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. Author manuscript; available in PMC: 2009 Sep 23.
Published in final edited form as: Circulation. 2009 Apr 20;119(17):2313–2322. doi: 10.1161/CIRCULATIONAHA.108.782458

Figure 2.

Figure 2

Nuclear NF-κB proteins of PASMCs specifically bind to the −254(C→G) SNP–generated NF-κB binding site. A, NF-κB activation demonstrated by translocation of p50 and p65 from the cytoplasm to the nucleus. PASMCs from normal subjects were stimulated with (+) or without (−) TNF-α (10 ng/mL). B, Electrophoretic mobility shift assay and competition analysis. TNF-α–stimulated nuclear extracts from PASMCs were incubated with biotin-labeled double-stranded oligonucleotides corresponding to the 24-bp sequences containing the wild-type −254C or mutant −254G probes. Nuclear extract/oligonucleotide complexes (C1 and C2) are indicated by arrows. C, Antibody-mediated supershift assay. NF-κB p50 and p65 antibodies identified complexes C1 as p50/p65 and C2 as p50/p50 NF-κB–containing complexes.