FIG. 6.

Effects of Ad5IκB and LY 294002 on IL-8 induction by TNF-α. Hc cells infected with Ad5IκB or Ad5LacZ were pretreated with or without 25 μM LY 294002 for 1 h prior to treatment with TNF-α (20 ng/ml). (A) Total RNA (20 μg) from Hc cells at 1 h after TNF-α administration was subjected to Northern blot analysis using the specific probe against IL-8 synthesized by RT-PCR. (B) Total cell lysate of Hc cells at 3 h after TNF-α administration was subjected to Western blotting with antibodies against IL-8 and actin. (C) At 5 h after TNF-α administration, the culture medium was harvested, concentrated, and subjected to Western blotting with antibody against IL-8. The results shown are representative of at least two independent experiments.