Additional file 1: Figure S1. Chemical structure of EE-15-one. The chemical
structure of 2-Ethyl-estra-1,3,5(10),15-tetraen-3-ol-17-one (EE-15-one)
showing the ketone group at position C17 which makes EE-15-one an
estrone derivative rather than an estradiol derivative along with an alkene
group at C15. Additionally, the chemical structures of ESE-15-one, EE-one,
EE-15-ol are depicted.
Additional file 2: Figure S2. EE-15-one related cell detachment in MDAMB-
231 cells is not due to induced cell death. Medium from MDA-MB-231
cells exposed to 5 μM EE-15-one was collected at the indicated times and
analysed for LDH activity. The graph shows the percentage LDH activity as
compared to the positive control. The average result of three independent
experiments was plotted with error bars representing s.e.m. *Indicates significant
differences between negative control medium and medium from
EE-15-one treated cells as calculated by student’s t-test at a P < 0.05.
Additional file 3: Video S1. EE-15-one exposure results in the disassembly
of the focal adhesions, severance of actin stress fibres connected
to focal adhesions. For timelapse imaging, MCF-7 cells were transfected
with LifeAct-mRFP (actin, red) and paxillin-GFP (focal adhesions, green).
The following day cells were prepared for imaging every 30 s on a Zeiss
LSM800 confocal microscope using a 63× magnification objective with
environmental control. Cells were incubated with 5 μM EE-15-one. Scale
bar, 20 μm.