Trizol reagent, SuperScript III Reverse Transcriptase and Lipofectamine 2000 were obtained from Invitrogen. Mouse antibody for beta actin and rabbit antibody for HIF 1alpha had been purchased from Genete . Human and mouse recombinant PlGF protein and an enzyme linked immuno sorbent assay kit have been obtained from R and D Techniques. A dual luciferase reporter assay technique was obtained from Promega. Hemato ylin and Eosin, Chromatin immuno precipitation Assay Kit, and EZ Zyme Chromatin Prep Kit were bought from Merck Millipore. An in situ cell Death Detection Kit and tremeGENE HP DNA Transfection Reagent have been obtained from Roche. The FITC Anne in V apoptosis detection Kit I was obtained from BD Biosciences. The JNK inhibitor, SP600125, was obtained from Enzo Daily life Science.

A SuperSensitive Polymer HRP IHC Detection Technique was obtained from Biogene . Animals This research Inhibitors,Modulators,Libraries conformed towards the Pointers for the Care and Use of Laboratory Animals published through the United states Nationwide Institutes of Well being. Each of the animal e periments were accepted through the Institutional Animal Care and Use Committee with the Laboratory Animal Center, College of Medication and Public Health of Nationwide Taiwan University. Eight week old male C57BL 6 wild type mice were bought through the Laboratory Animal Center, College of Medicine and College of Public Wellbeing, Nationwide Taiwan University. The PlGF knockout mice in B6 background were provided by Dr. Po Nien Tsao. Cell culture Human bronchial epithelial cells, BEAS 2B, have been cultured in F12 nutrient mi ture with 0. five ng ml recombinant epidermal growth element, 500 ng ml hydrocortisone, 0.

005 mg ml insulin, Inhibitors,Modulators,Libraries 0. 035 mg ml bovine pituitary e tract, 500 nM ethanolamine, 500 nM phosphoethanolamine, 0. 01 mg ml transferrin, 6. five ng ml three, three, five triiodothyronine, 500 ng ml epinephrine, 0. one ng ml retinoic acid, 10% FCS one hundred unit ml penicillin, and a hundred Entinostat ug ml streptomycin in the humidified 95% air 5% CO2 incubator at 37 Inhibitors,Modulators,Libraries C. Mouse primary form II alveolar epithelial cells and culture medium have been obtained from chi scientific. Principal ordinary human bronchial epithelial cells have been kindly provided by Dr. Reen Wu at University of California, Davis. Plasmids Human genomic DNA was e tracted from BEAS 2B by a Speedy gDNA MiniPrep kit. The 2.

0 kb human PlGF promoter Inhibitors,Modulators,Libraries region was amplified from human genomic DNA applying polymerase chain response performed with Hi Fi Taq DNA polymerase as follows two minutes at 94 C, then 15 sec at 94 C, thirty sec at 59 C, and 2 min and 30 sec at 72 C for 35 cycles. The amplified DNA fragments have been cloned into pGL3 vector as well as sequences had been confirmed by DNA sequence evaluation. The pGL3 with mouse PlGF promoter was as previously described. Enzyme linked immuno sorbent assay Cellular medium from BEAS 2B and AEC II, and BAL fluid from mice were analyzed by a PlGF ELISA kit according for the manufacturers directions.