Fig. 11

Breast cancer cells exposed to EO CM exhibit a reduced number of cells in S phase of cell cycle. MDA-MB-231 or MCF-7 cells were plated at 5 × 10³ cells in 4-well chamber slides in 50% growth media plus either 50% vehicle media (control) or 50% EO CM. Culture media was also supplemented with 0.5 μM 5-ethynyl-2-doxyuridine (EdU) for imaging using the Click-iT EdU Imaging Kit. Cells were maintained in media plus EdU for the entire length of the time course. On days 1, 2, 3, 4, and 5 post-treatment, cells were fixed with 4% paraformaldehyde and washed with PBS. For EdU imaging, cells were permeabilized, washed, then incubated with the Click-iT reaction cocktail. Cells were subsequently stained with DAPI, then visualized using fluorescence microscopy. *P < 0.01, **P < 0.005, ^#P < 0.0005, ^##P < 0.0001 +EO CM vs. +DMEM or +EMEM treatment. N = 3 biological replicates per condition, per time point