Bank vole were treated with 50 µg/kg bw; six doses every other day

MDA-MB-231 Cells were seeded into 3, 96-well plastic tissue culture dishes per line at 1,000 cells per well on day 0. On day 1, each plate was treated with DY131 (5 and 10 mkM) . Plates were re-dosed every 3 days and stained on days 3 or 4, 7 or 8, and 10 or 11. To stain, one plate per line was rinsed once with 1X Phosphate-Buffered Saline (PBS) and incubated with a solution of 0.5% w/v crystal violet (Sigma Aldrich) dissolved in 25% methanol: 75% water at 4°C for 10 minutes. Excess stain was removed and each plate was washed 5-6 times with deionized H2O and allowed to air dry completely. Stained cells were rehydrated in a 0.1M sodium citrate buffer dissolved in 50% ethanol: 50% water, then read on a plate reader at an absorbance of 550nm. Each experimental condition was performed in six replicate wells.