FIGURE 2.

RANTES induction is highly NF-κB-dependent. HeLa cells were pretreated with either Me₂SO or 5 μm BAY for 30 min prior to treatment with vehicle (PBS) or TNF (20 ng/ml) for 30 min. A and B, cells were fixed and incubated with anti-p65 (A) or anti-p50 antibodies (B). A secondary antibody with a 488 fluorophore was utilized, and cells were visualized by confocal microscopy. C, HeLa cells were treated with vehicle (Me₂SO) or BAY (5 μm) for 30 min prior to stimulation with PBS or TNF (20 ng/ml) for 24 h. RNA was extracted, and RANTES mRNA levels were assessed by qRT-PCR. The data represent means ± S.E. of three independent experiments. D, HeLa cells were treated with 20 nm Astar or p65 siRNA for 48 h and treated with vehicle (PBS) or TNF (20 ng/ml) for 24 h, and the levels of RANTES mRNA were assessed by qRT-PCR. The data represent means ± S.E. of three independent experiments. *, p < 0.05; **, p < 0.01.