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. 1997 Feb 3;185(3):481–490. doi: 10.1084/jem.185.3.481

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Activation of NF-κB is not inhibited by CrmA (A) or Bcl-2 (B). Electrophoretic mobility shift assay (EMSA) for the transcription factor NF-κB in MCF-7 cells expressing CrmA, Bcl-2, or vector is shown. Cells were seeded at 2 × 10⁶ cells/10-cm dish in RPMI media containing 10% FBS and rested overnight. Treatment with 2 nM TNF-α proceeded for 30 min after which the cells were trypsinized, nuclear extracts prepared, and EMSA performed using 10 μg of nuclear protein and a ³²P-labeled NF-κB oligonucleotide probe as described in Materials and Methods. Bands representing the specific NF-κB-DNA complex, a nonspecific band (n.s.), and the free probe are indicated.

Activation of NF-κB is not inhibited by CrmA (A) or Bcl-2 (B). Electrophoretic mobility shift assay (EMSA) for the transcription factor NF-κB in MCF-7 cells expressing CrmA, Bcl-2, or vector is shown. Cells were seeded at 2 × 10⁶ cells/10-cm dish in RPMI media containing 10% FBS and rested overnight. Treatment with 2 nM TNF-α proceeded for 30 min after which the cells were trypsinized, nuclear extracts prepared, and EMSA performed using 10 μg of nuclear protein and a ³²P-labeled NF-κB oligonucleotide probe as described in Materials and Methods. Bands representing the specific NF-κB-DNA complex, a nonspecific band (n.s.), and the free probe are indicated.