Anti mmp9 ab76003

The cells were lysed with RIPA buffer (Millipore, Billerica, MA, USA) containing protease inhibitor cocktail (Roche), and the lysates were quantified with a protein assay kit (Bio-Rad, Hercules, CA, USA). The cell lysates were separated using SDS-PAGE and then transferred to PVDF membranes. The proteins were identified while using appropriate antibodies. Anti-VGLL1 (10124-2-AP) was purchased from Proteintech (Rosemont, IL, USA). Anti-TEAD4 (ab58310) and anti-MMP9 (ab76003) were purchased from Abcam (Cambrige, MA, USA). Anti-GFP (NB600-308) was purchased from Novus Biologicals (Centennial, CO, USA). Anti-Myc (sc-789) and anti-HA (sc-805) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-GAPDH (LF-PA0212) was purchased from AbFrontier (Seoul, Korea). Anti-β-tubulin (2128), anti-pSer473-AKT (9271), anti-AKT (9272), anti-p-β-catenin (9561), and anti-β-catenin (9562) were purchased from Cell Signaling (Danvers, MA, USA). Anti-Flag (F1804) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The protein signal was detected while using an enhanced chemiluminescence kit (Millipore, Burlington, MA, USA).

SiRNAs (CCR2/p65/scrambled) were from Santa Cruz Biotechnology and siRNA transfection reagent Hiperfect were obtained from Qiagen. RT‐PCR reagents and enzymes were from Invitrogen. Monoclonal antibodies, anti‐human MMP‐1 (MAB901), anti‐human CCR2 (MAB180) were from R&D Systems, UK, anti‐MMP‐9 (ab76003) from Abcam, anti‐phospho‐PKC (A‐11) from Santa Cruz and anti‐actin (A5060) from Sigma‐Aldrich. Human MCP‐1 ELISA kit (DuoSet) and Go 6976 (PKC inhibitor) were from R&D Systems. All other substances and materials were from Sigma‐Aldrich, Invitrogen, Life Technologies, Eppendorf, Bio‐Rad.