FIGURE 5.

Salinomycin blocks TGFβ-induced myofibroblast contraction and migration. A, human fibroblasts were cultured in a collagen gel matrix as described in the text. After collagen gel formation, cells were treated with TGFβ (5 ng/ml) or TGFβ plus either salinomycin (SNC, 250 nm) or narasin (Nar, 250 nm) and allowed to contract for 72 h. Gel matrices were imaged at the 0, 24, 48, and 72 h time points. The gel area was quantified using ImageJ. TGFβ-induced contraction was set to 100%, and all other measurements were normalized to it. Representative images are shown, demonstrating that TGFβ readily induced myofibroblast contraction, whereas salinomycin and narasin inhibit contraction by more than 80% and 90%, respectively, 72 h post-TGFβ. *, p < 0.01 versus TGFβ-treated cells. B, human fibroblasts were cultured in a 12-well culture dish until they reached a confluent monolayer. A scratch wound was then induced in the cultures, and wells were washed to remove cells and debris. Cells were treated with TGFβ (5 ng/ml) or TGFβ plus salinomycin (250 nm) for 72 h to allow myofibroblast migration. Open areas were quantified using ImageJ software. Areas were normalized to time 0 (area = 1.0 at time 0). Images were captured at 0, 24, 48, and 72 h. After 72 h, TGFβ-treated cells had reduced the scratched area to less than 20% of the original area. Salinomycin treatment prevented myofibroblast migration 3- to 4-fold over TGFβ only. *, p < 0.01 versus TGFβ-treated cells. Experiments were repeated in triplicate, with two different strains and representative images shown.