The effects of inhibiting actomyosin dynamics and E-cadherin function on mESC organization on flat substrates from Physical confinement signals regulate the organization of stem cells in three dimensions

Embryonic stem cells were seeded onto a flat PDMS surface with inhibitory drugs (a) blebbistatin (2 µM)(n=24), (b) Y27632 (10µM)(n=20) and (c) SMIFH2 (10µM)(n=28) for a 48hr incubation period (Scale bar = 50 µm and applies to all). Contrary to cells grown in a confined microenvironment, inhibition of actin contractility, mechanotransduction signaling and actin polimerization respectively had no effect on aggregate morphology whilst on a two dimensional surface. (d) No significant difference was found with regards to the planar Isotropy (Ip, black) or globular isotropy (Ig, red) when we used any of the inhibitory conditions.