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. 2011 Nov 7;287(1):245–256. doi: 10.1074/jbc.M111.274613

FIGURE 3.

FIGURE 3.

Direct effect of 3-FC on p65-DNA binding. A, 3-FC directly affects NF-κB-DNA binding. Nuclear extracts (NE) were prepared from untreated cells or cells treated with TNF-α (0.1 nm), incubated with 100 μm 3-FC for the indicated times, and then analyzed for NF-κB activation by EMSA. B, DTT reverses the inhibitory effect of 3-FC on NF-κB-DNA binding. Nuclear extracts of TNF-α-treated KBM-5 cells were incubated with 100 μm 3-FC for 30 min in the absence or presence of 100 μm DTT and then assayed for NF-κB activation using EMSA. C, DTT reverses the inhibitory effect of 3-FC on the binding of reconstituted p65 to DNA. The p65−/− cells were transfected with p65 plasmid, and nuclear extracts were incubated with 100 μm 3-FC with or without DTT for 30 min and then assayed for p65 binding to DNA by EMSA. D, lack of effect of 3-FC on DNA binding of mutant p65 (p65 mt (C38S)). The p65−/− cells were transfected with wild-type and mutant p65 plasmids. Nuclear extracts of transfected cells were incubated with 3-FC for 30 min and then assayed for DNA binding by EMSA.