Fig. 3. ISO inhibits constitutive activation of NF-κB in breast cancer cells. (A, B) MDA-MB-231 cells were incubated with the indicated concentrations of ISO for 24 h or with 80 μM of ISO for indicated time. The nuclear extracts were analyzed for NF-κB-DNA binding activity; bars give the standard error (**p<0.01, ***p<0.001 vs control). (C) MDA-MB-231 cells were incubated with 80 μM of ISO for 24 h. The intracellular distribution of NF-κB was analyzed by immunofluorescence assay. After cells were fixed and permeabilized, NF-κB (green) was stained with rabbit anti-NF-κB followed by FITC-conjugated secondary antibodies and nuclei (blue) were stained with Hoechst 33342. The third panels display the merged images of the first and second panels.
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