Epothilone b

Cell viability was determined by CCK8 cytotoxicity assay. Briefly, cells were seeded in 96-well plates at 5000 cells/well and treated with at 37°C with Ispinesib (0, 30, 60, 120 or 240 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No.HY-50759), Paclitaxel (0, 10, 20, 40 or 80 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No. HY-B0015) and Epothilone-b (0, 40, 80, 160 or 320 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No. HY-17029) for 24 h, respectively. Cells were incubated with CCK8 (10 μl/well) for 2 h at 37°C and then spectrophotometrically quantified at 450 nm.

We performed a CCK-8 assay (Cellcook, Guangzhou, China, Cat No. CT01A) using the manufacturer’s guidelines to determine the viability of cells transfected with DDIT4-siRNA and siRNA-NC. We seeded these cells in the logarithmic growth phase into 96-well plates. In order to evaluate the effect on cell proliferation capacity, 10 μl CCK-8 reagent was added in all wells and incubated for 2 hours at 37°C at 0, 24, 48, 72 h after culturing. For the sensitivity of cells to the drug, cells were treated at 37˚C with Ispinesib (0, 20, 40, 80 or 100 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No.HY-50759), Cabazitaxel (0, 10, 20, 40 or 80 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No. HY-15459) and Epothilone-b (0, 40, 80, 160 or 320 nM, MedChem Express, Monmouth Junction, NJ, USA, Cat No. HY-17029) for 24 h, respectively. 10 μl CCK-8 reagent was added in all wells and incubated for 2 hours at 37°C. Finally, we measured the absorbance of each well at 450 nm using a microplate reader.