Additional file 1: of Anti-EMT properties of CoQ0 attributed to PI3K/AKT/NFKB/MMP-9 signaling pathway through ROS-mediated apoptosis

CoQ0-induced apoptosis in MDA-MB-231 cells. The cells were exposed to CoQ0 (5–15 μM for 24 h). (a-b) The TUNEL assay was performed to determine apoptotic DNA fragmentation. The green florescence indicates the number of TUNEL positive cells in the microscopic fields (400 × magnification) from three separate samples. The percentage of apoptotic cells was calculated by measuring the florescence intensity of treated cells using commercially available software. (c) Annexin V-FITC and PI staining was used to identify the early/late apoptosis or necrosis, and the data were analyzed using flow cytometry. The results in each quadrant are labeled and interpreted as follows: (Q1) PI positive, Annexin V-FITC-negative stained cells/necrosis. (Q2) PI positive, Annexin V-FITC-positive stained cells/late apoptosis. (Q3) Cells negative for both PI and Annexin V-FITC staining/normal live cells. (Q4) PI-negative, Annexin V-FITC-positive stained cells/early apoptosis. (d) Effects of CoQ0 on apoptotic-related proteins. Protein levels of mitochondria/cytosolic cytochrome c, caspases-9, caspase-3, and PARP, Bax, Bcl-2, and p53 were analyzed by Western blotting. The results are presented as the mean ± SD of three independent assays. ***p < 0.001 significant compared to control cells. (PPTX 51519 kb