Figure 3

Cooperative induction of EMT in NHBE cells by TGF-β1 and TWEAK. Confluent monolayers of NHBE cells were cultured for 48 h in the absence (control) or presence of TGF-β1, TNF-α, TWEAK, or TGF-β1 in combination with TNF-α or TWEAK, as indicated. The levels of E-cadherin (A) and N-cadherin (B) mRNA were analyzed by qRT-PCR. Expression levels were normalized to the housekeeping gene GAPDH and calculated as fold induction in comparison to the control. Whole cell lysates were immunoblotted for E-cadherin (C, upper) and N-cadherin (D, upper). All membranes were re-probed with anti-α-tublin antibody to confirm equal loading. The density of each band was normalized with α-tubulin and quantified by densitometry using NIH ImageJ software (C and D, lower). The capability of cell migration was assessed using a wound-healing assay (E). Data represent the means ± SD of three independent experiments. ^* p < 0.05 compared with the untreated culture. ^† p < 0.05 compared with TGF-β1 alone.