Effect of EMT induction on adhesion activity of A549 cells.

<p>(A and B) A ninety-six-well plate was coated with 2 µg/ml laminin-332 (Lm332), 5 µg/ml fibronectin (FN), and 2 µg/ml collagen I (Col I) at 4°C overnight, and then blocked with 1.2% BSA for 1 h at 37°C. A549 cells were incubated with 10 ng/ml TGF-ß in serum-free medium for 24 h. The TGF-ß-treated cells (TGF-ß) and untreated cells (Control) were suspended and inoculated onto the plate. After incubation for 30 min, phase contrast images were taken (A) and cell adhesion activity was measured (B). A scale bar indicates 50 µm. The relative numbers of attached cells were determined as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053209#s4" target="_blank">Materials and Methods</a>. Each bar indicates the mean ± SD of adherent cells in triplicate wells. (C) Electric time-lapse cell adhesion assay with E-plates. The plate was precoated with fibronectin (○, •) and collagen I (□, ▪) as shown above, and the adhesion of TGF-ß-treated cells (TGF-ß: •, ▪) and untreated cells (Control: ○, □) to the plate was monitored every 5 min. Cell index indicates arbitrary unit reflecting attachment and spreading of the cells on microelectrode array in the bottom of the wells. Other experimental conditions are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053209#s4" target="_blank">Materials and Methods</a>.