Figure 4.

Effect of MiTF on IL-8 in human cervical stromal cells. A, IL-8 mRNA and secreted protein were quantified in cells treated with control (CTL), MiTF-CX, mutant MiTF-CX (MiTF^mi), or MiTF-CX + MiTF^mi adenovirus. *, P ≤ 0.01 compared with CTL B, HEK293 (b and c) and cervical stromal cells (e and f) were transfected with wild-type MiTF-CX (WT; b and e) or MiTF-CX^mi (mi; c and f) and immunocytochemistry was conducted to localize MiTF protein using FITC or Cy3. MEL cells (a) served as positive controls for MiTF localization. MiTF immunostaining was absent in nontransfected cervical stromal cells in culture (d). C, Cervical stromal cells or HEK293 cells were treated with increasing amounts of MiTF-CX^mi with or without increasing levels of wild-type MiTF-CX. Total viral titers in plaque-forming units were equivalent in all conditions. After 48 h, IL-8 mRNA levels were quantified using qPCR. Data represent mean of triplicate determinations on each sample. D, Immunoblot analysis of MiTF in cytosolic (Cyt) and nuclear (N) extracts from cervical stromal cells treated with control virus (Ctl), MiTF-CX (10⁷ pfu, MiTF), MiTF-CX^mi (10⁸ pfu, MiTF^mi), or both. Molecular weight markers are noted on the right.