Pcdna3.1 runx2

The rno-miR-137-3p mimics (sequence: 5′-UUAUUGCUUAAGAAUACGCGUAG-3′), mimics negative control (NC; sequence: 5′-UUGUACUACACAAAAGUACUG-3′), rno-miR-137-3p inhibitor (sequence: 5′-CUACGCGUAUUCUUAAGCAAUAA-3′) and inhibitor NC (sequence: 5′-CAGUACUUUUGUGUAGUACAA-3′) were synthesized by GenePharma (Shanghai, China). The constructs, including pmirGLO-wt-Runx2, pmirGLO-mt-Runx2, pmirGLO-wt-CXCL12, pmirGLO-mt-CXCL12, pmirGLO-Runx2-PC, pmirGLO-CXCL12-PC, pcDNA3.1-Runx2, pcDNA3.1-CXCL12 and lentiviral vector pEZX-MR03 were also obtained from GenePharma.
The antibodies used for western blotting in our study were: anti-Runx2 (Abcam, Cambridge, UK, 1:1000), anti-CXCL12 (Abcam, 1:1000), anti-β-actin (Sigma-Aldrich, St Louis, MO, USA, 1:2000), and rabbit secondary antibody (Biosynthesis Biotechnology, Beijing, China, 1:5000). The antibodies used for immunohistochemistry were: anti-Runx2 (Abcam, 1:200), type I collagen (COL I; Abcam, 1:200), VEGF (Abcam, 1:200), and SDF-1α (Abcam, 1:150).

The transfection of PC3 cells was achieved using 50 nM of miR-373 Mimics (Mimics) or miR-NC Mimics (NC Mimics) using Lipofectamine 2000 (Invitrogen, USA) for 48 h. pcDNA3.1 and pcDNA3.1-RUNX2 were synthesized and provided by GenePharma (Shanghai, China). The transfection of the cells was achieved using either pcDNA3.1-RUNX2 or pcDNA3.1 based on the manufacturer's protocols that use Lipofectamine 2000.