Figure 4

Effect of pre-exposure to DON on the capability of A431 to respond to mechanical stimulation. (a) Images are representative of A431 cells after incubation in static (+STATIC 24 h) or mechanically stimulated environment (+STRETCHED 24 h, 0.5 Hz, 15% deformation) showing cell nuclei (blue), lysosomes (red), tubulin (green) and plasma membrane (white) after live cell imaging. (b) Quantification of the average cell number/optical fields (o.f.); n ≥ 10 randomly acquired optical fields from at least 3 independent cell preparations. *Indicates at 0.05 level significant increase in comparison to STATIC incubation. ^δδδIndicates at 0.001 level significant decrease in comparison to STATIC incubation (Mann-Whitney test). (c) Measurement of cell viability. WST-1 data are expressed as % of cell growth between PRE-POST incubation (n ≥ 3 membranes for each experimental condition). ^##Indicates at 0.01 significant decrease in comparison to STATIC and STRETCHED controls (DMSO 1:1000; ANOVA). (d) Average cell area; n ≥ 45 cells randomly chosen from 3 independent experiments (**and ***indicates at 0.01 and 0.001 level significant increase in comparison to STATIC incubation with Student’s t-test). Data are expressed as mean ± S.E. and describe controls (DMSO 1:1000, white graphs) and cells pre-incubated with DON (grey graphs) at static incubations (solid bars) and mechanically stimulated incubations (striped bars).