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10.1248/bpb.31.2255 http://scihub22266oqcxt.onion/10.1248/bpb.31.2255 19043209!ä!19043209 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 286.79999999999995 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Biol+Pharm+Bull 2008 ; 31 (12): 2255-9 Nephropedia Template TP {{tp|p=19043209|t=2008. Establishment of a cell-based assay to screen regulators of the hypoxia-inducible factor-1-dependent vascular endothelial growth factor promoter |pdf=|usr=}}{{19043209}} gab.com Text Establishment of a cell-based assay to screen regulators of the hypoxia-inducible factor-1-dependent vascular endothelial growth factor promoter JO: Biol Pharm Bull http://www.kidney.de/pget.php?&tw=1&pmid=19043209 #FOAvip In this study, we established a drug screening system based on transcriptional regulation of vascular endothelial growth factor (VEGF) under hypoxia-inducible factor-1alpha control. We cloned the neomycin-resistance gene into the plasmid GL (pGL)3-promoter vector to generate the pGL3-promoter-neo vector. The 3 copies of the 47-bp fragment that contained the hypoxia response element of VEGF were synthesized and inserted in front of the minimal promoter of the pGL3-promoter-neo vector to generate p3HRE-luc-neo. The recombinant reporter gene vectors were transfected into EAhy926 cells, and stable cell lines were obtained. The positive cell line was selected for its ability to express luciferase in response to hypoxia. We demonstrated that CoCl(2) significantly enhances luciferase activity in a concentration-dependent fashion. We then optimized the cell density and incubation time under hypoxia which were used to screen. The assay exhibited a low background and was an ideal model for high-throughput screening for human VEGF regulators. Twit Text FOAVip Establishment of a cell-based assay to screen regulators of the hypoxia-inducible factor-1-dependent vascular endothelial growth factor promoter ????Biol Pharm Bull http://www.kidney.de/pget.php?&tw=1&pmid=19043209 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=19043209 #FOAvip English Wikipedia <ref>{{Cite pmid|19043209}}</ref> Establishment of a cell-based assay to screen regulators of the hypoxia-inducible factor-1-dependent vascular endothelial growth factor promoter #MMPMID19043209 Ji DB; Zhu HB; Ye J; Li CL Biol Pharm Bull 2008[Dec]; 31 (12): 2255-9 PMID19043209show ga In this study, we established a drug screening system based on transcriptional regulation of vascular endothelial growth factor (VEGF) under hypoxia-inducible factor-1alpha control. We cloned the neomycin-resistance gene into the plasmid GL (pGL)3-promoter vector to generate the pGL3-promoter-neo vector. The 3 copies of the 47-bp fragment that contained the hypoxia response element of VEGF were synthesized and inserted in front of the minimal promoter of the pGL3-promoter-neo vector to generate p3HRE-luc-neo. The recombinant reporter gene vectors were transfected into EAhy926 cells, and stable cell lines were obtained. The positive cell line was selected for its ability to express luciferase in response to hypoxia. We demonstrated that CoCl(2) significantly enhances luciferase activity in a concentration-dependent fashion. We then optimized the cell density and incubation time under hypoxia which were used to screen. The assay exhibited a low background and was an ideal model for high-throughput screening for human VEGF regulators. |Cell Line[MESH] |Cell Survival/drug effects[MESH] |Cobalt/pharmacology[MESH] |DNA/biosynthesis/genetics[MESH] |Drug Evaluation, Preclinical[MESH] |Drug Resistance[MESH] |Endothelial Cells/drug effects/metabolism[MESH] |Humans[MESH] |Hypoxia-Inducible Factor 1/*physiology[MESH] |Luciferases/biosynthesis/genetics[MESH] |Neomycin/pharmacology[MESH] |Neovascularization, Physiologic/drug effects[MESH] |Plasmids/genetics[MESH] |Promoter Regions, Genetic/*genetics[MESH] |Protein Synthesis Inhibitors/pharmacology[MESH] |Reproducibility of Results[MESH] |Tetrazolium Salts[MESH] |Thiazoles[MESH] |Transfection[MESH]Establishment of a cell-based assay to screen regulators of the hypoxi(epmc).pdf DeepDyve Pubget Overpricing