Figure 2.

Over-expression of ANT1 decreased the activity of NF-κB. A. ANT1 transfection reduced nuclear translocation of NF-κB by ANT1 over-expression. MDA-MB-231 cells were transfected with pcDNA-ANT1 and 12 hours later cells were blocked with 3% fetal bovine serum in 1% BSA/PBS for 30 min, washed with PBS three times, and incubated overnight with anti-NF-κB antibody. After washing three times, cells were incubated with anti-IgG-FITC or DAPI for 30 min. Images were obtained using a LSM510. B. ANT1 transfection reduced the nuclear translocation of NF-κB by ANT1 over-expression. Cells were transfected with pcDNA-ANT1; 12 hours later, total or nuclear extracts were prepared and Western blotted using anti-NF-κB or anti-α-tubulin antibodies. Immunoblots were visualized using an enhanced chemiluminescence detection system. C. ANT1 transfection reduced the transcriptional activity of NF-κB induced by ANT1 over-expression. Cells were co-transfected with pcDNA-ANT1 and NF-κB-luciferase reporter construct for 12 hours and then lysed. Lysate luciferase activities were determined using a luminometer.