The miR-491 (pre-miR-491) expression vector and related control vector were constructed by synthetic oligonucleotides and cloned into the pcDNA6.2-GW/EmGFP vector (Invitrogen) between the EcoRI and HindIII sites as previously described.37 (link), 38 (link) Using bioinformatic analyses, we identified a fragment of Wnt3a as a miR-491-5p target. The 3′-UTR of the human Wnt3a mRNA was constructed by synthetic oligonucleotides and cloned into the pmir GLODual-Luciferase miRNA Target Expression Vector (Promega, Madison, WI, USA) between the SacI and XhoI sites as previously described.37 (link), 38 (link) A miR-491-5p inhibitor, an inhibitor negative control, and siRNAs targeting Wnt3a were chemically synthesized (GenePharma, Shanghai, China). Wnt3a was cloned into the GV230 vector (Genechem, Shanghai, China) between the XhoI and KpnI sites. Foxi1 was cloned into PEGFP-C1 between the HindIII and BamHI sites (GENEWIZ, Suzhou, China). A 127 bp sequence containing the wild-type or mutant Foxi1-binding site was cloned into the pGL3 promoter vector between the KpnI and XhoI sites (GENEWIZ). All the sequence information is given in Supplementary Table S2 .
Pegfp c1
Manufactured by Genewiz
Sourced in China
The PEGFP-C1 is a plasmid vector that expresses the enhanced green fluorescent protein (EGFP) under the control of the CMV promoter. It is used for the expression and localization of EGFP-tagged fusion proteins in mammalian cells.
Automatically generated - may contain errors
Lab products found in correlation
Pcdna6.2 gw emgfp vector, by Thermo Fisher Scientific (1 mentions)
Pmirglo dual luciferase mirna target expression vector, by Promega (1 mentions)
Gv230 vector, by Genechem (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Thermo Fisher Scientific (1 mentions)
Luciferase assay system, by Promega (1 mentions)
Ferric chloride hexahydrate, by Shanghai Chemical Reagent (1 mentions)
Ammonium hydroxide, by Shanghai Chemical Reagent (1 mentions)
Dimethyl sulfoxide (dmso), by Shanghai Chemical Reagent (1 mentions)
2 protocols using pegfp c1
1
Functional Validation of miR-491-5p Targets
2
Carboxylated Carbon Nanotubes for Biomedicine
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Carboxyl functionalized multi-wall carbon nanotubes (CNTs-COOH) were obtained from the Chinese Academy of Sciences Chengdu Organic Chemicals Co., LTD. The purity is 98 wt% and contains 1.23 wt% carboxyl groups. The length is 0.5-2 μm and the outer diameter is 20-30 nm. 1.8k PEI(Mw=1.8 kDa)and 25k BPEI(Mw=25 kDa) were obtained from Sigma-Aldrich. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS) were purchased from Aladdin Co. (Shanghai, China). Acrylic acid, cyclohexane, 1,6-hexanediol, triethylamine, dimethyl sulfoxide (DMSO), ammonium hydroxide, sodium bicarbonate, ferrous chloride tetrahydrate, ferric chloride hexahydrate, and hydroquinone were purchased from Shanghai Chemical Reagent Company and were used as received.
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) was purchased from Invitrogen. Dulbecco's Modified Eagle's Medium (DMEM) and penicillin-streptomycin were purchased from Gibco. Fetal bovine serum (FBS) was purchased from Hyclone. Luciferase Assay System was purchased from Promega.
Hela and COS-7 cells were incubated in DMEM containing 10% FBS at 37 °C and humidified air/5% CO 2 . The plasmids DNA of pEGFP-C1 and pGL3-Luc were purchased from Genewiz (Suzhou, China).
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) was purchased from Invitrogen. Dulbecco's Modified Eagle's Medium (DMEM) and penicillin-streptomycin were purchased from Gibco. Fetal bovine serum (FBS) was purchased from Hyclone. Luciferase Assay System was purchased from Promega.
Hela and COS-7 cells were incubated in DMEM containing 10% FBS at 37 °C and humidified air/5% CO 2 . The plasmids DNA of pEGFP-C1 and pGL3-Luc were purchased from Genewiz (Suzhou, China).
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