Figure 1

DcR3 promotes cellular migration in RCC. (A) Immunoblot analysis of whole-cell lysates showing DcR3 protein levels in different human RCC cell lines and human embryonic kidney 293-T cells. (B) Immunoblot analysis of precipitated protein from supernatant showing DcR3 protein secretion in different human RCC cell lines and human embryonic kidney 293-T cells. Ponceau staining is used to demonstrate equal protein loading. (C,D) Immunoblot analysis of whole-cell lysates (left) and precipitated protein from supernatant (right) of 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 control vector (neo) (D). (E) Scratch motility assay of ACHN and 769-P cells transfected with two different DcR3-specific siRNAs or a non-specific siRNA (scram). Bars indicate the percentage of cell migration in relation to cells transfected with a non-specific siRNA after 24 h (mean ± SEM; n=3; *p<0.05, **p<0.01; T-test). Representative images are shown in Additional file 1: Figure S1D. (F) Scratch motility assay of ACHN and 769-P cells stably overexpressing DcR3 or an empty control vector (neo). Migration was measured over a time course of 12 h (ACHN) or 24 h (769-P). Bars indicate the percentage of cell migration in relation to cells stably transfected with an empty control vector (mean ± SEM; n=3; *p<0.05, **p<0.01, T-test). Representative images are shown in Additional file 1: Figure S1E. (G) Scratch motility assay of ACHN and 769-P cells transfected with two different DcR3-specific siRNAs or a non-specific siRNA (scram) and incubated with either DcR3-containing or control supernatant (SN) of stable transfectants. Migration was measured over a time course of 24 h. Bars indicate the percentage of cell migration in relation to cells transfected with a non–specific siRNA and treated with control supernatant (mean ± SEM; n=3; *p<0.05, **p<0.01; T-test).