Rabbit anti human mmp9

Western blots were prepared as previously described [8 (link)]. Antibodies used were: rabbit anti-human ANXA4 (1:200, Santa Cruz), mouse anti-human ANXA4 (1:1000, Abcam), mouse anti-human Lewis y antigen(1:500, Abcam), rabbit anti-human NF- KB p50 (1:200, Santa Cruz), mouse anti-human NF-kB p50 (1:200, Santa Cruz), rabbit anti-human integrin α5 (1:500, Proteintech, Wuhan, China), rabbit anti-human integrin β1 (1:500, Proteintech), rabbit anti-human integrin β5 (1:200, Beijing Biosynthesis Biotechnology Co., Ltd., Beijing, China), rabbit anti-human MMP2 (1:500, Proteintech), rabbit anti-human MMP9 (1:500, Proteintech), rabbit anti-human LC-3 (1:1000, Abcam), and rabbit anti-human Bcl-1 (1:1000, Cell Signaling Technology, Danvers, MA, USA).

Western blots were performed as described pre-viously [24 (link)]. Briefly, the total protein extract of each tissue sample or cell line was dissolved in a lysis buffer (25 mM Tris–HCl pH 7.4,1 % Triton X-100, 150 mM NaCl, 5 % EDTA, 10 mM NaF, 1 mM phenylmethylsulfonyl fluoride, and 10 mg of aprotinin and leupeptin), separated on 10% SDS-PAGE gels, transferred to PVDF membranes (Millipore, USA) and quantitated using a BCA protein assay kit. Equal amounts of protein (60 μg) were analyzed by immunoblotting. The antibodies used during Western blots included rabbit anti-human ERα from Proteintech, rabbit anti-human ERβ (1:500) from Proteintech, mouse anti-human MMP-1(1:400) from Santa Cruz, mouse anti-human MMP-2(1:400) from Santa Cruz, rabbit anti-human MMP-9 (1:500) from Proteintech, rabbit anti-human p38MAPK(1:500)/p-p38MAPK(1:500) and rabbit anti-human tAkt (1:500)/pAkt(1:500) from Bioworld Technology. Rabbit anti-human GAPDH (1:5000) from Santa Cruz was used as an internal control for protein loading and analysis. The protein bands were visualized using enhanced chemiluminescence (ECL) kit (Pierce, USA).