Rabbit anti human ki67

Immunohistochemical analyses of MP expression, Ki₆₇ antigen, and microvessel density (MVD, CD31) were carried out with rabbit anti-MP, rabbit anti-human Ki₆₇ (Millipore Corporation, Billerica, MA, U.S.), and rabbit anti-mouse CD31 antibodies (Abcam PLC, Boston, MA, U.S.) using the labeled streptavidin-biotin method as previously described19 (link). Apoptotic cells in tumor tissues were detected in paraffin sections using a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling DeadEnd Fluorometric (TUNEL) system assay kit (Promega, Madison, WI, U.S.) according to the manufacturer’s instructions19 (link). The quantification of MVD and TUNEL-positive cells was assessed according to previous reports19 (link)57 (link)60 (link).
The sections were also stained with hematoxylin and eosin (H&E) for histomorphometric analysis. All tissues of three mice in each group were randomly examined with H&E/IHC. All sections were observed or counted by two investigators or pathologists in a blinded fashion.

For ISH detection, deparaffinized tissue sections were hybridized with 50 nM LNA-modified and digoxin-labeled miR-320a oligonucleotide probe (Exiqon, Vedbaek, Denmark; Supplementary Table 4) for 1 h at 55°C, incubated with 5μg/ml anti-digoxin- Rhodamine antibody (Roche Applied Science, Indianapolis, IN, USA) overnight at 4°C, and stained with DAPI in the dark for 15 min at room temperature (RT). The IHC staining was performed according to the standard ABC protocol with the antibodies of mouse anti-human SND1 (Abcam, Cambridge, MA, USA), rabbit anti-human β-catenin (CST, Boston, MA, USA) and rabbit anti-human Ki-67 (Millipore, Billerica, MA, USA). The labeling index (LI) is presented as the percentage of positive cell number to total cell number.