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1994). The constant �� (mN/mm2) has units of force that in slowly contracting muscle (i.e., smooth muscle) is independent of the load, while ��/�� (lref/s) is a constant with units of velocity. selleckchem This index of the RSi reflects all internal factors that increasingly reduce shortening velocity of unloaded muscle during contraction, that is, those intracellular components, such as cytoskeletal proteins, that are compressed during shortening. In lung sections from the same lobes described above for CD4+ staining, we then evaluated by IHC the presence in the airways of the cytoskeletal protein vimentin, which has been shown to play an find more important role in tissue mechanical stability (Goldman et al. 1996; Eckes et al. 1998; Wang and Stamenovic 2000). After antigen retrieval with citrate buffer, pH 6.0, and blocking of nonspecific binding with 10% normal horse serum, slides were incubated overnight at 4��C with monoclonal mouse antivimentin antibody, which was ready to use (clone V9, AbD Serotec). As secondary antibody we used biotinylated horse anti�\mouse, 1:200 dilution for 30 min at room temperature. Detection was performed with ABC complex, followed by DAB peroxidase reaction and slides were counterstained with methyl green. Drugs and chemicals Indomethacin were purchased from Sigma Chemical Co.; pentobarbital sodium from Abbot Laboratories, Chicago, IL; 10% neutral Vatalanib (PTK787) 2HCl buffered formalin from Trend Scientific Inc., St. Paul, MN. Data analysis Data are expressed as mean �� SEM, except when differently indicated. Data were compared using either Student's t�\test or analysis of variance (ANOVA). The least significant difference (LSD) post hoc analysis was used to find which groups were responsible for differences revealed by ANOVA. Statistical analysis was carried out using SigmaStat (V. 3.5, Systat, San Jose, CA). Values of P