The distal femoral bone marrow cavity was exposed to prepare decalcified bone slides

Samples were decalcified at room temperature in 15% EDTA for 5 weeks. After decalcification, the sample was placed in 70% alcohol and paraffin embedded. Four micrometer paraffin slides were prepared on glass slides coated BYL719 PI3K inhibitor with egg white-glycerol, or polylysine, then dried for one hour at 60uC and stored at 4uC for future use. Immunohistochemical analysis of bone marrow microcirculation factor VIII-related antigen and peroxisome proliferator-activated receptor c were performed following the manufacturers’ instructions. Rabbit antihuman vWF polyclonal antibody Carfilzomib PR-171 Proteasome inhibitor| was purchased from CHEMICON International, Inc, USA, and PPARc mouse monoclonal antibody was purchased from Santa Cruz Biotechnology, Inc, USA. Briefly, the endogenous peroxidase activity of slides were blocked by 3% hydrogen peroxide and nonspecific binding was blocked with 10% goat serum.