Mature human miR-26b (5’-UUCAAGUAAUUCAGGAUAGGU-3’), miR-26b inhibitor (anti-miR-26b, 5’-ACCUAUCCUGAAUUACUUGAA-3’) and control microRNA sequence (miR-C, 5’-GUUCUAGUACAAUAUUAGGAG-3’) were purchased from RiboBio Co. Ltd (Guangzhou, China). For overexpression of ATF2, open reading frame region of human ATF2 was amplified and linked to pcDNA3.1 eukaryotic expression plasmid (Invitrogen, USA). For knockdown of ATF2, ATF2 siRNA was purchased from Genepharma Company (Shanghai, China). For transfection, 2 μg/ml ATF2 plasmid, 50 pmol/ml miR-26b, anti-miR-26b, miR-C and ATF2 siRNA were introduced into the Hep-2 and Hep-2/R cells by using lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.
Mir c
Manufactured by Thermo Fisher Scientific
Sourced in United States
The MiR-C is a compact and modular autonomous mobile robot designed for safe and efficient transportation of materials within indoor environments. It is capable of navigating through crowded areas and tight spaces, while avoiding obstacles and optimizing its route. The MiR-C is suitable for a variety of applications that require the movement of goods or materials within a facility.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Atf2 sirna, by GenePharma (1 mentions)
Pcdna3.1 eukaryotic expression plasmid, by Thermo Fisher Scientific (1 mentions)
Pcdna3.1 eukaryotic expression vector, by Thermo Fisher Scientific (1 mentions)
Mir c, by RiboBio (1 mentions)
Esirna1, by Merck Group (1 mentions)
Pre mir mirna precursor molecules, by Thermo Fisher Scientific (1 mentions)
Mir 99a, by Thermo Fisher Scientific (1 mentions)
Pmirglo3, by Promega (1 mentions)
Cyanine3 (cy3), by Thermo Fisher Scientific (1 mentions)
Pmirglo plasmid, by Promega (1 mentions)
4 protocols using mir c
1
Modulation of ATF2 by miR-26b in Hep-2 Cells
2
Overexpression of XIAP in BCSCs
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MiR-489 mimics (5′-GUGACAUCACAUAUACGGCAGC-3′) and random RNA oligonucleotides (miR-C, 5′-ACUGAAUACUGCGCAAACGUGC-3′) were purchased from RiboBio Co. Ltd. (Guangzhou, China).XIAP open reading frame was amplified and then linked to the pcDNA3.1 eukaryotic expression vector (Invitrogen) to overexpress the gene of XIAP in BCSCs. For transfection,cells were cultured in 6-well plates overnight followed by transfecting with miR-489 mimics (50 pmol/ml), miR-C (50 pmol/ml) and XIAP plasmid (2 μg/ml) by using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s protocols. After transfection, cells were collected for the following experiments.
3
Transfection of Luciferase, siRNA, and miRNA
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Lipofectamine 2000 (Life Technologies) was used to transfect luciferase plasmids, silencing RNA or miRNA molecules. Pre-designed MISSION siRNAs specific to human c-Myc (esiRNA1, Sigma-Aldrich) were used to knock down c-Myc expression. MISSION siRNAs are endoribonuclease-prepared siRNA pools comprised of a heterogeneous mixture of siRNAs that all target the same mRNA sequence. Cells were transfected with siRNAs for 48 h prior to harvest for protein or RNA extraction. To overexpress miR-92a, an engineered miR-92a mimetic molecule (Ambion’s Pre-mir MiRNA Precursor Molecules) was used to transfect HCT116 cells according to the manufacturer’s protocol. miR-C (Ambion) was used as a control.
4
Luciferase Assay for miR-99a Regulation
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The luciferase experiments were performed in HEK293T cells. HEK293T cells were seeded at 1 × 104 cells per well in a 96-well plate and were transfected the following day with Lipofectamine with the following molecules: the synthetic miRNA precursor miR-99a (ID: AM17100; Thermo Fisher Scientific), and negative control miR-C (ID: AM17110; Ambion), Cy3 (ID: AM17020; Thermo Fisher Scientific) and the pmirGLO plasmid (Promega Corporation, Madison, WI, USA) containing the luciferase reporter and also the Renilla gene (control) versus pmirGLO3′-UTR-E2F2 or pmirGLO3′-UTR-EMR2. The transfection efficiency was ∼95%, and luciferase activity was measured 48 h after transfection with the dual luciferase reporter assay as previously described.39 (link) In each case, the miR-99a concentrations were measured by titrating the miR-99a with each pmirGLO3′-UTR mRNA construct to establish a dose–response relationship between 10 and 80 nM (data not shown). Results were conducted in triplicate in at least three independent experiments.
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