Figure 4. TG2-knocked-down or IL-6-knocked-down ovarian cancer cells revealed reduced epithelial-to-mesenchymal transition and in vitro migration. (A) Intracellular activity and expression of TG2 in the lung tissues from MDAH-2774 cells and cultivated peritoneal mass. Enzyme activity was estimated by biotinylated pentylamine (BP) incorporation probed with HRP-conjugated streptavidin (SA) in Western blot analysis. (B) Morphology of MDAH-2774 cells and cultivated peritoneal mass. Peritoneal masses from MDAH-2774-bearing mice were removed and digested with collagenase D and DNase I. Cells were suspended in 30% Percoll, layered onto the top of a 70% Percoll gradient, and centrifuged. The top layer, an enriched population of epithelial and fibroblast cells, was retained. Human cells were selected using MACS separation with antihuman MHC class I antibody (anti-HLAI). (C) Expression of TG2, fibronectin, and vimentin in MDAH-2774 cells and peritoneal mass was determined by Western blot analysis. (D) Expression of matrix metalloproteinases in MDAH-2774 cells and peritoneal mass were determined by reverse transcription-polymerase chain reaction. (E) Migration of MDAH-2774 cells into the wound areas created by scratching was induced by IL-6 (10 ng/mL) for 20 h. These results represent one of three independent experiments.