Paclitaxel ptx

A total of 2000 cells/well were plated on 96-well plates. After 24 hours, the cells were exposed to cytotoxic agents at various concentrations. Hydrogen peroxide (H 2 O 2 ) (Wako, Osaka, Japan) was added to the culture medium for 10 minutes. The medium was then replaced with fresh medium, and cell viability was measured after 48 hours. Iron (III) chloride (WAKO) was added to the culture medium, and cell viability was measured after 48 hours. The anticancer drugs, cisplatin (CDDP) (Sigma-Aldrich) and paclitaxel (PTX) (Wako) were diluted with dimethylformamide (DMFA) and added to the culture medium, and cell viability was measured after 72 hours. Each result was obtained from 3 independent experiments with 8 replicates.

The PI3K inhibitor, wortmannin (Sigma-Aldrich, St Louis, MO, USA), was used to inhibit the PI3K pathway. The MEK 1/2 inhibitor, U0126 (Sigma-Aldrich), was used to inhibit the MAPK pathway. The inhibitory effect of wortmannin (1 μM) and U0126 (10 μM) was confirmed by western blotting (Supplementary Figure 1D). The SIRT1 selective inhibitor, EX527 (Merck Millipore, Billerica, MA, USA) was used at functional concentrations as described previously. 19 The anticancer drugs, cisplatin (CDDP) (Sigma-Aldrich) and paclitaxel (PTX) (Wako, Osaka, Japan), diluted with 5% dimethylformamide (DMFA) and saline were added to the culture medium at various concentrations, and cell viability was measured after 72 h.
Cell viability in the assay for proliferation or anticancer drug resistance was evaluated using the WST-1 reagent (Roche Diagnostics, Basel, Switzerland) according to the manufacturer's instructions. Briefly, cells were seeded onto 96-well plates. After culturing the cells under various conditions, the WST-1 reagent was added to the medium. After 2.5 h, A450 wavelength was measured using the microplate reader, SYNERGY HT (BioTek, Winooski, VT, USA). Each result was obtained from three independent experiments with 16 replicates.