Peroxidase conjugated goat anti rabbit antibody

Quercetin (Sigma, USA) was prepared into a 200 mM stock using dimethyl sulfoxide (DMSO) and stored at −20°C. The working concentration of 200 μM Quercetin was made in a complete medium, and a range of concentrations of Quercetin (10, 20, 50, 100, 150 and 200 μM) was utilized in cell viability assay. Cisplatin (Sigma, USA) and paclitaxel (MedChemExpress, USA) were stored at −20°C, while 5-fluorouracil (Solarbio, China) and doxorubicin (Solarbio, China) were stored at 4°C. Primary antibodies for MMP2 (monoclonal rabbit anti-human antibody; 1:1000), ezrin (polyclonal rabbit anti-human; 1:1000) and GAPDH (monoclonal rabbit anti-human antibody; 1:1000) were purchased from Cell Signaling Technology, and P-Gp (monoclonal rabbit anti-human antibody; 1:1000) and METTL3 (monoclonal rabbit anti-human antibody; 1:1000) were purchased from Abcam (1:1000, USA); and the second antibody, peroxidase-conjugated goat anti-rabbit antibody, was obtained from Biosharp (1:5000, USA). Diaminobenzidine (DAB) Substrate Kit was purchased from Zhongshan Goldenbridge (China).

Quercetin dihydrate (Sigma, USA) was dissolved in dimethyl sulfoxide (DMSO; Corning, USA) and further diluted with medium to reach 0, 20, 40, 50, 80, 100, 150, 160, and 200 μM before treatment. The following primary antibodies were used in the present study: rabbit anti-human Fascin antibody (1:1000; Abcam, USA), rabbit anti-human Erzin antibody (1:1000; Cell Signaling Technology, USA), mouse anti-human MMP-2 antibody (1:1000; Cell Signaling Technology, USA), rabbit anti-human MMP-9 antibody (1:1000; Cell Signaling Technology, USA), mouse anti-human β-actin antibody (1:3000; Abcam, USA), rabbit anti-human GAPDH antibody (1:3000; Abcam, USA), mouse anti-human vimentin antibody (1:400; Proteintech, USA), and mouse anti-human cytokeratin antibody (1:400; Proteintech, USA). The secondary antibodies used in this study were listed as follows: peroxidase-conjugated goat anti-rabbit antibody (1:5000; Biosharp, China) and anti-mouse antibody (1:5000; Biosharp, China).