Figure S5 Low dose anti-tumor agent treatment de- creased the expression of uPA and uPAR. Following 24 h drug treatments, total RNA extracted from osteosarcoma cell lines were analyzed by real-time PCR. Treatment with cisplatin, methotrexate, etoposide or doxorubicin decreased uPA and uPAR expression in 143B and Saos-2 cells (P,0.05). The comparative Ct (DDCt) method was used to determine fold change in expression using GAPDH or ACTB. Experiments were performed in triplicate with similar results [error bars represent mean (SD)]. (TIF) Figure S6 Chemotherapy prevents expression of uPA and uPAR by down-regulation of EGR1. Twenty four hours after 4 mg doxorubicin treatment, RNA was extracted from tumour in nude mice xenograft model. Real-time PCR revealed that chemotherapy decreased uPA and uPAR expression in vivo (A) (P,0.05). To examined whether EGR1 affects the expression of uPA and uPAR in vivo. 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