Fig. 3.

Triptonide markedly reduces Twist1 protein levels in TNBC cells and in the tumor tissues of xenografted mice. MDA-MB-231 and MDA-MB-468 cells were treated with triptonide (TN) for 72 h at the concentrations of 0–10 nM. Twist1 protein levels were detected using Western blotting (A, B), and the bands were scanned and statistically analyzed (C). Additionally, the cells were treated with 10 nM TN for 0, 12, 24, 36, 48, and 72 h, respectively, Twist1 protein levels were measured (D, F) and statistically analyzed (E, G). Twsist1 protein levels in MDA-MB-231 and MDA-MB-468 cells were also detected using immunofluorescent staining and confocal microscopy (H, I, × 2400). Furthermore, Twist1 protein levels in the lung tissues of xenografted mice were detected by immunohistochemistry staining (J, × 1000) and statistically analyzed (K). The results represent three independent experiments. *P < 0 .05, **P < 0.01