Figure 3. Capsanthin decreases cellular EZH2 and suppresses the binding of p21 promoter by EZH2 in TNBC cells. (A) Proteins were extracted from synchronized MDA-MB-231 cells at various times after treatment with 10 μM capsanthin for 0, 3, 6, 9, 12 or 24 h and then cell cycle–related proteins were analyzed with western blotting. (B) Synchronized MDA-MB-231 cells were treated with 10 μM capsanthin for 0, 3, 6, or 9 h, and qPCR was carried out to assess the cellular level of EZH2 mRNA. *P < 0.05, **P < 0.01, ***P < 0.001. (C) MDA-MB-231 cells were treated as described for panel A, and then proteins were extracted from the cells and subjected to western blotting for EZH2 and H3K27me3. (D) MDA-MB-231 and BT-549 with capsanthin downregulates EZH2 level and upregulates the p21 promoter. Capsanthin-treated cells were subjected to qChIP to assess the activation of the p21 promoter. *P < 0.05. Cap, capsanthin.