Adenoviral vectors carrying CTRP3 small hairpin RNAs (shRNAs) designed to knock down CTRP3 in adipocytes were generated by Hanbio (Shanghai, China). Scrambled shRNA was used as a control. Briefly, adipocytes were incubated in DMEM/F12 and were infected with adenoviral vectors at a multiplicity of infection (MOI) of 100 for 4 h. To knock down Nrf2 in H9c2 cells, we purchased siNrf2 and negative-control siRNA from Guangzhou RiboBio Co., Ltd. H9c2 cells were transfected with siNrf2 using Lipofectamine RNAiMAX (Invitrogen) for 24 h. Transfection efficiency was confirmed by western blotting. AntagomiR-200a and a miRNA inhibitor control, which were obtained from Guangzhou RiboBio Co., Ltd, were used to silence miR-200a in H9c2 cells.
Antagomir 200a
Manufactured by RiboBio
AntagomiR-200a is a synthetic oligonucleotide designed to inhibit the function of microRNA-200a. It acts as an antagonist to miR-200a, a regulatory RNA molecule involved in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Sinrf2, by RiboBio (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Scramble control, by OriGene (1 mentions)
Pmir report vector, by Thermo Fisher Scientific (1 mentions)
Siport neofx transfection agent, by Thermo Fisher Scientific (1 mentions)
Klf4 sirna, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
2 protocols using antagomir 200a
1
Adenoviral Knockdown of CTRP3 in Adipocytes
2
Keap1 3'-UTR Regulation by miR-200a
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pGL3-ARE and pcDNA3-HA-Nrf2 were generous gifts from Professor Xiaoming Yang. The 3′-UTR of Keap1 was amplified using the following primers: 5′-TCATACTAGTGGCACTTTTGTTTCTTGGGC-3′ and 5′-GCATTAAGCTTCAGGGTGAAAGACACTAG-3′ and cloned into pMiR-Report vector (Ambion) digested with HindIII and Spe I. We also generated three bases mutation in the predicated target site for miR-200a by using a QuickChange site-specific mutagenesis kit (Stratagene). All constructs were sequenced in Sangon Company. Transfection of plasmids was performed in 70%–80% confluent cells using Lipofectamine 2000 Reagent (Invitrogen) according to manufacturer's protocol.
MSA was purchased from Sigma–Aldrich (Sigma–Aldrich Inc.). Pre-miR miR-200a precursor and Pre-miR negative control were purchased from Ambion. Antagomir-200a was synthesized from Ribobio. KLF4 siRNA and scramble control were purchased from OriGene (OriGene Technologies). Transfections of Pre-miR miR-200a precursor, Pre-miR negative control, KLF4 siRNA and scramble control were performed by using siPORT NeoFX Transfection Agent (Life Technologies) according to manufacturer's protocol.
MSA was purchased from Sigma–Aldrich (Sigma–Aldrich Inc.). Pre-miR miR-200a precursor and Pre-miR negative control were purchased from Ambion. Antagomir-200a was synthesized from Ribobio. KLF4 siRNA and scramble control were purchased from OriGene (OriGene Technologies). Transfections of Pre-miR miR-200a precursor, Pre-miR negative control, KLF4 siRNA and scramble control were performed by using siPORT NeoFX Transfection Agent (Life Technologies) according to manufacturer's protocol.
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