Wst 8

Cell growth and viability were measured using cell proliferation and cytotoxicity reagent WST-8 (Roche Biochemicals, Mannheim, Germany). Briefly, the protocol was as follows: 786-O, ACHN, Caki-1 cells (5 × 10³ per well) were cultured in a 96-well plate. After 12 h (time for cells to attach to the plate surface), cells were treated with different concentrations of CPS (0–400 μM) or pretreated with CPZ (2 μM) for 2 h and then treated with CPS, ten wells each group for statistics. At the harvest time 10 μl of CCK8 was added into each well and after one hour’s incubation cell viability was determined by measuring the absorbance of the converted dye at 490 nm. The experiments were triplicate.

The cell proliferation reagent WST-8 (Roche, Germany) was measured. Cell growth: 10 μL of cell counting kit-8 (CCK8) was added to each well at the time of harvest after plating cells in 96-well microtiter plates (Corning, NY) at 1.0 × 10³/well, according to the manufacturer’s instructions. Cellular viability was determined by measuring the absorbance of the converted dye at 450 nm 2 h after adding CCK8.

Cell growth was measured using the cell proliferation reagent WST-8 (Roche Biochemicals, Mannheim, Germany). After plating cells in 96-well microtiter plates (Corning Costar, Corning, NY) at 1.0× 10³ /well, 10 μL of CCK8 was added to each well at the time of harvest, according to the manufacturer's instructions. Two hours after adding CCK8, cellular viability was determined by measuring the absorbance of the converted dye at 450 nm.

Cell viability assays were performed using WST-8 (Roche Diagnostics, Mannheim, Germany) according to the manufacturer’s instructions^{18 (link)}. Cells were cultured in 24-well plates. Twenty-four hours after treatment with the indicated concentrations of A419259 (Tocris Bioscience), SKI-1 (Abcam, Cambridge, UK), PP2 (Cayman Chemical, MI, USA), saracatinib (Santa Cruz) or bosutinib (abcam), cells were incubated with the WST-8 substrate for 2–3 h, after which the absorbance of the wells was measured at 450 nm using a plate reader (2300 EnSpire^TM, PerkinElmer, Yokohama, Japan). The number of live cells was determined using a Countess cell counter (Invitrogen) after cultures were stained with trypan blue. Toxicity assays were performed using the Cytotoxicity Detection Kit PLUS (Roche Diagnostics, Indianapolis, IN). Twenty-four hours after treating the cells, 100 μL of the medium were removed from the plate and used in the assay. The medium was incubated with the substrate for 15 min and spectrophotometrically assayed at 490 nm using a plate reader. To activate the Src vector, we used MLR1023 (#4582) (Tocris Bioscience, Bristol, UK).

CCK8 Assay was carried out using the protocol described previously [7 (link), 18 (link), 21 (link), 47 (link)]. Briefly, cell growth was measured using the cell proliferation reagent WST-8 (Roche Biochemicals, Mannheim, Germany). After plating cells in 96-well microtiter plates (Corning Costar, Corning, NY) at 1.0× 10³ /well, 10 μL of CCK8 was added to each well at the time of harvest, according to the manufacturer's instructions. One hour after adding CCK8, cellular viability was determined by measuring the absorbance of the converted dye at 450 nm.

Cell proliferation was measured using the cell proliferation reagent WST-8 (Roche Biochemicals). After plating cells in 96-well microtiter plates (Corning Costar) at 1.0 × 10³/well, 10 μl of CCK-8 was added to each well at the time of harvest, according to the manufacturer's instructions. 2 h after adding CCK-8, cellular viability was determined by measuring the absorbance of the converted dye at 450 nm.