(A) MDA-MB-231 cells were treated with DMSO or Compound 1s (10 μM) and
then stimulated with sorbitol, as indicated. Cell lysates were analyzed by
western blot for phosphoERK5 (pERK5) and total ERK5. Data are representative of
two independent experiments. (B) A cell migration “scratch”
assay was employed using MDA-MB-231 cells. After scratching the cell monolayers,
cells were treated at 0 h with 1% DMSO (solvent), an inactive analog of
compound 1, the MEK5 inhibitor BIX02188 or Compound 1s (10 or 1 μM).
Photographs show the scratch at 0 and 18 hr as indicated. White areas are cell
monolayers, while black areas are zones without cells. Each condition was
performed in duplicate or singlet wells and data are representative of four
independent experiments.