The growth of parental and eribulin-resistant MDA-MB-231, Hs578T, and MDA-MB-157 cells pre-treated with DMSO and VOR (5 µM) or RICO (5 µM) for 48 h was measured by performing a WST assay (Wako Chemicals, Osaka, Japan). Brie y, 1 × 10 5 cells/well were seeded in 6-well plate and cultured for 24 h. Thereafter, a medium change was performed with DMSO, VOR, or RICO. After incubation for another 48 h, 4 × 10 3 cells/well were seeded in 96-well tissue culture plates in 100 µl of medium without VOR or RICO. After each indicated period, the absorbance was measured after adding WST solution. Each experiment was independently performed and repeated at least three times.
Small Interfering RNA (siRNA) Transfection ON-TARGETplus siRNA targeting ZEB1 (M-006564) and the negative control (D-001810) were purchased from GE Healthcare (Buckinghamshire, UK). Transfection of each siRNA (10 nM) was performed using Lipofectamine RNAi-MAX (Thermo Fisher Scienti c, Waltham, MA) following the manufacturer's instructions. Twenty-four hours after transfection, the proteins were extracted and 4 × 10 3 cells/well were cultured in 96-well tissue culture plates and incubated for 72 h after adding stepwise-diluted VOR or RICO.
Finally, the absorbance was measured after adding WST solution, as described previously.
Small Interfering RNA (siRNA) Transfection ON-TARGETplus siRNA targeting ZEB1 (M-006564) and the negative control (D-001810) were purchased from GE Healthcare (Buckinghamshire, UK). Transfection of each siRNA (10 nM) was performed using Lipofectamine RNAi-MAX (Thermo Fisher Scienti c, Waltham, MA) following the manufacturer's instructions. Twenty-four hours after transfection, the proteins were extracted and 4 × 10 3 cells/well were cultured in 96-well tissue culture plates and incubated for 72 h after adding stepwise-diluted VOR or RICO.
Finally, the absorbance was measured after adding WST solution, as described previously.