Cell viability assay was carried out as mentioned previously [38 (link)]. Briefly, HNSCC cells were seeded into 96 well plates at a density of 4 × 103 cells/well.
Cell lines were left overnight to attach, then treated with decreasing concentrations of paclitaxel in duplicates. Following this, 72 h after treatment, medium was removed and 50 μL PBS containing 1 mg/mL 3-(4,5-dimethylthiaziazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was added to each well and cells were incubated for 1 h at 37 °C. After the incubation MTT solution was removed and tetrazolium crystals were dissolved in isopropanol containing 10% (V/V) Triton X-100 and 1% (V/V) 0.1 N HCl. Absorbance was measured at 570 nm and 690 nm with a Synergy multimode reader (BioTek, Budapest, Hungary). The 690 nm data was subtracted from the 570 nm for each well. Absolute IC50 values were calculated by non-linear regression using Graph Pad Prism 5 software (GraphPad Software, San Diego, CA, USA). Each experiment was repeated at least three times.
Cell lines were left overnight to attach, then treated with decreasing concentrations of paclitaxel in duplicates. Following this, 72 h after treatment, medium was removed and 50 μL PBS containing 1 mg/mL 3-(4,5-dimethylthiaziazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was added to each well and cells were incubated for 1 h at 37 °C. After the incubation MTT solution was removed and tetrazolium crystals were dissolved in isopropanol containing 10% (V/V) Triton X-100 and 1% (V/V) 0.1 N HCl. Absorbance was measured at 570 nm and 690 nm with a Synergy multimode reader (BioTek, Budapest, Hungary). The 690 nm data was subtracted from the 570 nm for each well. Absolute IC50 values were calculated by non-linear regression using Graph Pad Prism 5 software (GraphPad Software, San Diego, CA, USA). Each experiment was repeated at least three times.
Free full text: Click here
