Monoclonal antibody for α smooth muscle actin

Sprague–Dawley rats were purchased from Charles River Japan (Hino, Japan). Dulbecco's modified Eagle's medium (DMEM) and DMEM/F-12 without phenol red, fetal bovine serum (FBS), trypsin-ethylenediaminetetraacetic acid (EDTA), and penicillin-streptomycin were purchased from GIBCO BRL (Grand Island, NY, USA). Human plasminogen and bovine fibrinogen were obtained from Enzyme Research Laboratories, Inc. (Swamsea, UK). Human urokinase and bovine thrombin were purchased from Calbiochem (Darmstadt, Germany). Monoclonal antibody for α-smooth muscle actin was purchased from DAKO (Glostrup, Denmark). 2′,7′-dichlorofluorescin diacetate (DCF-DA) was obtained from Molecular Probes (Eugene, OR, USA) and was dissolved in dimethyl sulfoxide (DMSO). 1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride (MPP) and 4-[2-phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]-pyrimidin-3-yl]phenol (PHTPP) were purchased from Tocris Bioscience (Bristol, UK). LysoPC and all other chemicals were obtained from Sigma Chemical Co. (St. Louis, MO, USA). E₂ was dissolved in ethanol (EtOH), whereas ICI 182,780, MPP, and PHTPP were dissolved in DMSO. All of the other chemicals were dissolved in water.

Sprague–Dawley rats were purchased from Charles River Japan (Hino, Japan). Dulbecco's modified Eagle's medium (DMEM) and Ham's F-12 medium (DMEM/F-12) without phenol red, fetal bovine serum (FBS), trypsin-ethylenediaminetetraacetic acid (EDTA), and penicillin-streptomycin were obtained from GIBCO BRL (Grand Island, NY, USA). 2′,7′-dichlorofluorescein diacetate (DCF-DA) was purchased from (Calbiochem, Darmstadt, Germany). LysoPC, 17β-E₂, ICI 182,780, and all other chemicals were from Sigma Chemical Co. (St. Louis, MO, USA). Monoclonal antibody for α-smooth muscle actin was obtained from DAKO (Glostrup, Denmark), anti-β-actin antibody was from Sigma Chemical Co., and antibodies against Bax, Bcl-2, precursors of caspase-9 and -8, active form of caspase-3, and cytochrome c oxidase subunit IV were bought from Cell Signaling Technology, Inc. (Beverly, MA, USA). DCF-DA and ICI 182,780 were dissolved in dimethyl sulfoxide (DMSO) and 17β-E₂ in ethanol (EtOH). All the other chemicals were resolved in distilled water.