Analysis of CXCL12 expression in prostate cancer cell lines stably overexpressing SLUG. (A, B) Analysis of RNA transcripts of CXCL12 by qPCR (left panel) and RT-PCR (right panel). RNA was extracted from PC3 (A) or DU-145 (B) prostate cancer cell lines infected with pMIGR1-Slug or pMIGR1 (vector) retroviruses, and used to synthesize cDNA. Transcript level of CXCL12 and SLUG was analyzed by qPCR (left panel) and RT-PCR (right panel). GAPDH was included as a loading control. CXCL12 was significantly upregulated by SLUG overexpression in PC3 (A) and DU145 (B). (C) ELISA analysis of CXC12 protein in PC3 cell line. Conditioned cell culture medium was collected from the cell culture environment of PC3 cells infected with pMig (vector) or pMig-Slug retroviruses and centrifuged to remove cell lysates, and then used for ELISA.