Figure 1.

Generation of IL-1α-overexpressing MCF-7 cells. A: NF-κB DNA-binding activity in MCF-7 cells transfected with pcDNA3 or IL-1α expression vector. Individual G418-resistant colonies were examined for NF-κB DNA-binding activity by electrophoretic mobility shift assay. DNA binding of the general transcription factor SP-1 is also shown. B: Induction of NF-κB in HLF-1 by the CM from pcDNA3 and IL-1α-overexpressing clones. HLF-1 cells were incubated with CM from indicated cell lines for 1 hour. NF-κB and SP-1 DNA-binding activities were measured as described above. CM from ∼90% confluent cells was collected after overnight incubation in serum-free media. C: Induction of IL-8 in HLF-1 cells by CM from various cell types. HLF-1 was incubated with CM for 4 hours and IL-8 expression was measured by Northern blotting of total RNA.