Smad4 Knockdown and Overexpression in Murine and Human Cancer Cell Lines

CT26 cells, an undifferentiated murine adenocarcinoma-derived cell line from Balb/c mouse, and human lymph-node metastasis-derived cell line SW620 were maintained in RPMI containing 7% FBS. Smad4 knockdown stable single cell clones were generated from CT26 cells by transfecting lentiviral shRNA construct targeting Smad4 (Sigma, St Louis, MO, USA, NM_008540) using puromycin (5 μg ml⁻¹) selection. SW620 stable clones expressing Smad4 were established as described previously (Zhang et al, 2010 (link)). One parental, two vector control clones, and three transfected single clones from each cell line were used for the following experiments. Female Balb/c mice (8 weeks old) and athymic nude mice (6 weeks old) were used for the experiments. All of animal works were performed in accordance with IACUC and state and federal guidelines for the humane treatment and care of laboratory animals.