Fig. 5. Cells expressing a truncated ADAMTS12 lacking thrombospondin domains (ADAMTS-TS12) undergo an EMT following HGF stimulation. (A) Domain organization of ADAMTS-TS12. The position of the HisTag epitope is indicated. (B) Expression analysis of ADAMTS-TS12 by RT-PCR and western blot using an antibody against the HisTag. In both cases, the cMDCK clone was used as a control. Truncated ADAMTS12 is located in the conditioned medium of cells that stably express this form of the enzyme. In RT-PCR, amplification of the mRNA encoding -actin was used to ascertain RNA integrity and ensure equal loading. (C) Scattering assay (2D), tubulogenesis assay (3D) and immunostaining of E-cadherin and vimentin of the ADAMTS-TS12-expressing cells in the absence (– HGF) or presence (+ HGF) of hepatocyte growth factor. Bars, 30 µm (left, centre); 10 µm (right).