Figure 3

DcR3 modulates cell adhesion to ECM component fibronectin. (A,B) Adhesion assay of ACHN and 769-P cells transfected with two different DcR3-specific siRNAs or a non-specific siRNA (scram) (A) or stably overexpressing DcR3 or an empty vector control (neo) (B). Bars indicate the percentage of cell adhesion compared to control cells. Adherent cells were counted in five representative fields (mean ± SEM; n=3, *p<0.05, **p<0.01; ***p<0.001; T-test). Representative images are shown (magnification: 100×). (C,D) Quantitative real-time-PCR assaying relative expression levels of DcR3 and invasion-relevant genes in ACHN and 769-P cells transfected with two different DcR3-specific siRNAs or a non-specific siRNA (scram) (C) or stably overexpressing DcR3 or an empty vector control (neo) (D). Expression data were normalized to internal 18S rRNA expression. Bars indicate fold changes of RNA level in relation to control cells (mean ± SEM; n=3; *p<0.05; **p<0.01; T-test). (E) Immunoblot analysis of whole-cell lysates showing uPA protein level in ACHN and 769-P cells transfected with two different DcR3-specific siRNAs, a non-specific siRNA (scram) or stably overexpressing DcR3 or empty vector control (neo). (F) Immunoblot analysis of whole-cell lysates showing ITGA4 protein level in ACHN and 769-P cells stably overexpressing DcR3 or an empty vector control (neo).