Figure-S6 from <i>ASS1</i> as a Novel Tumor Suppressor Gene in Myxofibrosarcomas: Aberrant Loss via Epigenetic DNA Methylation Confers Aggressive Phenotypes, Negative Prognostic Impact, and Therapeutic Relevance

Abstract

<p>Figure-S6 PDF file 78K, Figure-S6 ASS1 knockdown promotes cell proliferation, viability, migration, and invasion in LPS510 cells that endogenously express ASS1. (A) Among the 4 liposarcoma cell lines tested, endogenous ASS1 mRNA and protein are only readily detected in LPS510 cells by quantitative RT-PCR (upper) and Western blotting assays (lower), respectively. CCD966SK fibroblasts are run in parallel as a positive control. (B) Stable silencing of ASS1 gene with shASS1 in LPS510 cells show significantly downregulated expression at the mRNA and protein levels as detected by real-time RT-PCR (upper) and Western blotting (lower) assays, respectively. The shLacZ plasmid, POLR2A transcript, and GADPH protein are utilized as controls in RNA interference, quantitative RT-PCR, and Western blotting assays, respectively. (C) Results of Brdu (left upper), XTT (right upper), transwell migration (left lower), and matrigel invasion (right lower) assays reveal that the functions of cell proliferation, cell viability, cell motility, and cell invasiveness are significantly impaired in shLacZ control (all p��0.05) as compared to the LPS510 cells stably silenced against ASS1 expression. All functional experiments are performed in triplicate and results expressed as mean �} SD</p>