Figure 5.

NO₂-OA inhibits TNFα-induced TNBC cell migration and invasion. A, experimental schemes and representative images of crystal violet-stained migrating MDA-MB-231 or MDA-MB-468 cells. Cells (1 × 10⁵) were placed in the upper chamber with serum-free medium under the indicated treatment conditions. Migrating cells were photographed using a light microscope at ×100. B and C, quantitation of migrated cells from Fig. 4A was performed by solubilization of crystal violet and spectrophotometric analysis at A₅₇₃ nm. The percentage of migrating cells in each treatment group was compared with numbers of migrating cells in the absence of TNFα stimulation (Serum Ctrl). *, p < 0.05 versus in the absence of TNFα stimulation; **, p < 0.05 versus TNFα alone. D, to test the impact of NO₂-OA on TNBC cell invasion, MDA-MB-468 cells were incubated in serum-free medium containing 20 ng/ml TNFα combined with NO₂-OA (5 μm), NO₂-SA (5 μm), or JSH-23 (10 μm), and then invasion was determined by the extents of cell migration through the Matrigel matrix toward a 5% FBS chemoattractant for 24 h. The percentage of invading cells in each treatment was relative to the number of migrating cells in the absence of TNFα stimulation. *, p < 0.05 versus TNFα alone n.s., not significant. Significance was determined by one-way analysis of variance followed by Tukey post hoc test. All data are mean ± S.D. (error bars).