FIGURE 2.

SDF-1α induces nuclear translocation of NF-κB in HNSCC. TB2-T1 and TCA8113 cells were serum-starved and treated with 50 ng/ml SDF-1α for various times. Nuclear proteins (10 μg) from treated and untreated samples were isolated and incubated with [γ-³²P]CTP-labeled NF-κB probe and DNA binding buffer for 30 min at room temperature. Electrophoretic mobility shift assay was performed, and samples were separated on a 5% polyacrylamide gel. Protein-DNA complexes were visualized by autoradiography. Supershift lane represents nuclear extracts of cells treated with SDF-1α for 30 min and then incubated with p65 antibody for 30 min at room temperature, followed by addition of NF-κB probe. Oct-1 nuclear protein served as loading control using ³²P-labeled Oct-1 DNA probe.