Figure 5

Quantitative analysis of formation and inhibition of CCID in LEC monolayers by MCF-7 spheroids formation. LECs were seeded into 24-well plates and allowed to grow for 2 days until confluence when LECs were stained with cytotracker green. (A) MCF-7spheroids, which were treated with different concentrations (solvent, 1, 5, 10, 15, and 25 μ M) of Bay11-7082 for 0.5 h at 37°C, were transferred on top of LECs. (B) Either LECs or MCF-7 spheroids were treated with the Bay11-7082 for 0.5 h, which was entirely washed off before both cell types were co-cultivated. The 3D-MCF-7 spheroids/LEC monolayer co-cultures were incubated for 4 h at 37°C. The size of CCIDs, which were formed by MCF-7 spheroids in the LEC monolayer in this time period, was measured using a Zeiss Axiovert microscope and Axiovision software. In the solvent treated controls, the CCID sizes in LEC monolayers were set 100%. For each condition, the gap area of at least 12 spheroids was measured. Error bars indicate standard error of the mean. Asterisks show significant differences in the inhibition of CCID formation compared with control (^*P<0.05).