MiR-20b mimics, scrambled miR-20b, CREB siRNA, and CREB control siRNA were obtained from Genepharma (Shanghai, China). The miRNA sequences used were as follows: miRNA-20b mimics, sense: 5’-CAA AGU GCU CAU AGU GCA GGU AG-3’, antisense: 5’-ACC UGC ACU AUG AGC ACU UUG UU-3’; scrambled miR-20b, sense: 5’-UUC UCC GAA CGU GUC ACG UTT-3’, antisense: 5’-ACG UGA CAC GUU CGG AGA ATT-3’; CREB siRNA, sense: 5’-GUC UCC ACA AGU CCA AAC ATT-3’, antisense: 5’-UGU UUG GAC UUG UGG AGA CTT-3’; and CREB control siRNA, sense: 5’-UUC UCC GAA CGU GUC ACG UTT-3’, antisense: 5’-ACG UGA CAC GUU CGG AGA ATT-3’. PCR forward and reverse primers were synthesized by Sangon Biotech (Shanghai, China). The primer sequences used were: GAPDH primer, sense: 5’-GGC AAA TTC AAC GGC ACA-3’, antisense: 5’-TCC ACG ACA TAC TCA GCA CC-3’; CREB primer, sense: 5’-TGC CAC ATT AGC CCA GGT A-3’, antisense: 5’-GGG AGG ACG CCA TAA CAA CT-3’; miR-20b primer, sense: 5’-ATG CCA AAG TGC TCA TAG TG-3’, antisense: 5’-GTG CAG GGT CCG AGG T-3’; miR-20b RT primer: 5’-GTC GTA TCC AGT GCA GGG TCC GAG GTA TTC GCA CTG GAT ACG ACC TAC CT-3’; U6 primer, sense: 5’-CTCGCT TCG GCA GCA CA-3’, antisense: 5’-AAC GCT TCA CGA ATT TGC GT-3’; and U6 RT primer: 5’-AAC GCT TCA CGA ATT TGC GT-3’.
Creb sirna
Manufactured by GenePharma
Sourced in China
CREB siRNA is a laboratory reagent designed to target and suppress the expression of the CREB (cAMP Response Element Binding Protein) gene. It functions as a tool for researchers to investigate the role of CREB in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions)
Pparγ sirna, by GenePharma (1 mentions)
Streptozocin stz, by Merck Group (1 mentions)
Kn 93, by Merck Group (1 mentions)
Anti vegf, by Proteintech (1 mentions)
Anti pparγ, by Proteintech (1 mentions)
Anti camkii, by Cell Signaling Technology (1 mentions)
Anti phospho camkii, by Cell Signaling Technology (1 mentions)
Anti gfap, by Cell Signaling Technology (1 mentions)
Gw9662, by Merck Group (1 mentions)
Anti creb, by Cell Signaling Technology (1 mentions)
Anti phospho creb, by Cell Signaling Technology (1 mentions)
Anti β actin, by Proteintech (1 mentions)
Fibronectin, by ScienCell (1 mentions)
Huvecs, by GenePharma (1 mentions)
Huvecs, by Charles River Laboratories (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Huvecs, by ScienCell (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Endothelial cell growth supplement (ecgs), by ScienCell (1 mentions)
3 protocols using creb sirna
1
miR-20b, CREB Regulation Protocol
2
Alpinia zerumbet Attenuates Diabetic Neuropathy
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The Fructus Alpiniae zerumbet (FAZ) was collected from Zhenfeng County, Guizhou Province, China. Streptozocin (STZ) was purchased from Sigma (St Louis, MO, USA). RGZ, GW9662 and KN93 were obtained from Sigma (St Louis, MO, USA) and Selleckchem (Shanghai, China), respectively. Commercial kits, measuring insulin, VEGF and blood glucose, were obtained from Elabscience Co. Ltd. (Shanghai, China), Xin Bo Sheng (ERC103, China), Yuanye company (Shanghai, China), respectively. The primary anti-bodies used in this study including anti-GFAP, anti-Phospho-CREB, anti-CREB, anti-Phospho-CaMK II, and anti-CaMK II were supplied by Cell Signaling Technology (Danvers, MA, USA); anti-PPAR-γ, anti-VEGF and anti-β-actin were purchased from Proteintech (Chicago, USA). All reagents used for qRT-PCR were obtained from Takara Bio. (Dalian, China). PPAR-γ siRNA and CREB siRNA were supplied by GenePharma (Shanghai, China).
3
Manipulating HUVEC Signaling Pathways
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HUVECs were purchased from ScienCell (Santiago, MN, United States; Lot Number 28433), and cultured in endothelial cell medium (ECM, ScienCell) supplemented with 1% endothelial cell growth supplement (ECGS, ScienCell) and 5% FBS. The culture surfaces were pre-coated by fibronectin (ScienCell) which was diluted in PBS (1:100). Cells from the fourth to sixth generations were used for the experiments. At 80%–90% confluence, HUVECs were transfected with control, Gsα, or CREB siRNA (GenePharma, Shanghai, China) using Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific). According to the manufacturer’s instructions, the culture medium was replaced with Opti-MEM (Gibco) and the RNA-lipid complexes were added to cells followed by incubating for 6 h at 37°C and replacing with normal medium. After 48 h, cells were collected for analysis. For the virus-mediated gene transduction, HUVECs were infected with adenovirus-expressing Gsα or lentivirus-expressing CREB (Vigenebio, Jinan, China) for 48 h followed by analysis. H89 and forskolin were purchased from Abcam.
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