Transfection was done on astrocytes, in 80% confluent T25 flasks using RNAi MAX (Invitrogen) as per the manufacturer's protocol. 62.5 nM of Syn‐hsa‐miR‐92b‐5p miScript miRNA mimic (Qiagen, Cat# MSY0004792) was used for mimic transfection. All‐Stars Negative Control siRNA (Qiagen, Cat# SI03650318) was used as mimic control. miRNA inhibition was done using 62.5 nM of the miRCURY LNA miRNA inhibitor against has‐miR‐92b‐5p (Qiagen, Cat# YI04104160‐ADAFr), and miRCURY LNA miRNA inhibitor control (Qiagen, Cat# YI00199006‐ADA) was used as the inhibitor control. Transfections were done using Opti‐MEM media (Gibco, Cat# 31985–070) for 5 h and replaced with complete MEM. Cells were processed after 48 h of transfection for RNA and protein analysis. Effective doses of mimic and inhibitor miRNA were standardized via qPCR and protein studies.
Mircury lna mirna inhibitor control
Manufactured by Qiagen
Sourced in Germany
The MiRCURY LNA miRNA Inhibitor Control is a synthetic, chemically modified oligonucleotide designed to inhibit the function of a specific microRNA (miRNA) in biological samples. It serves as a negative control for experiments involving miRNA inhibition.
Automatically generated - may contain errors
Lab products found in correlation
Syn hsa mir 92b 5p miscript mirna mimic, by Qiagen (3 mentions)
Mircury lna mirna inhibitor against has mir 92b 5p, by Qiagen (3 mentions)
Rnaimax, by Thermo Fisher Scientific (3 mentions)
Opti mem media, by Thermo Fisher Scientific (3 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Exo fect exosome transfection kit, by System Biosciences (1 mentions)
5 protocols using mircury lna mirna inhibitor control
1
Astrocyte miR-92b-5p Modulation
2
Plasmid Transfection and miRNA Inhibition
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pCMV-β-gal plasmid was obtained from Clontech Lab (Mountain View, CA, USA). hPromB8-LUC plasmid contains the firefly luciferase cDNA under the control of a -3000/+523 human HSPB8 promoter region (18 (link)); pCI-hHSPB8-wild-type (wt) codes for the human HSPB8 protein (26 (link)). pHSPB8-mut has been obtained in our laboratory by exchanging the ApaI/SalI coding fragment with the mutated sequence obtained from Eurofins Genomics. pEGFP-G93A-SOD1 expresses the green fluorescent protein (GFP)-tagged mutant G93A SOD1 (17 (link)). pcDNA3.1 (Life Technologies, #V790-20) plasmid was used to normalize the amount of transfected plasmid DNA. All plasmids were transfected in MCF-7 and MDA-MB-213 cell lines as previously described (21 (link)). NSC-34 cells were transfected as previously described (25 (link)). The hsa-miR-574-5p miRCURY LNA miRNA Inhibitor (Qiagen) was used to inhibit miR-574-5p activity, and the miRCURY LNA miRNA Inhibitor Control (Qiagen) was used as a control. Both miRNAs were transfected at the final concentration of 50nM according to the manufacturer's instructions.
3
Exosome-Mediated microRNA Modulation
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hMSC-EV transfection was performed using the Exo-FectTM Exosome Transfection Kit (EXFT20A-1, System Biosciences) with stabilized miR-antagomirs or a pair of miR-antagomirs (miRCURY LNA inhibitors for microRNAs hsa-miR-29c-3p, miR-21-5p, hsa-miR-129-5p, hsa-miR-92a-3p; miRCURY LNA miRNA Inhibitor Control, Qiagen). The transfected EVs were dissolved in fresh DMEM-LG up to the appropriate concentration, and in vitro or in vivo models were used.
4
miR-92b-5p Transfection in Astroglia
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Transfection was done on astroglia, in 80% con uent T25 asks using RNAi MAX (Invitrogen, USA) as per the manufacturer's protocol. 62.5 nM of Syn-hsa-miR-92b-5p miScript miRNA mimic (Qiagen, Germany) was used for mimic transfection. All-Stars Negative Control siRNA (Qiagen, Germany) was used as mimic control. miRNA inhibition was done using 62.5 nM of miRCURY LNA miRNA inhibitor against has-miR-92b-5p (Qiagen, Germany) and miRCURY LNA miRNA Inhibitor Control (Qiagen, Germany) was used as the inhibitor control. Transfections were done using Opti-MEM media (Invitrogen, USA) for 5 h and replaced with complete MEM. Cells were processed after 48 hours of transfection for RNA and Protein analysis. Effective doses of mimic and inhibitor miRNA were standardized via qPCR and protein studies.
5
Transfection of Astrocytes with miR-92b-5p
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Transfection was done on astrocytes, in 80% con uent T25 asks using RNAi MAX (Invitrogen, USA) as per the manufacturer's protocol. 62.5 nM of Syn-hsa-miR-92b-5p miScript miRNA mimic (Qiagen, Germany) was used for mimic transfection. All-Stars Negative Control siRNA (Qiagen, Germany) was used as mimic control. miRNA inhibition was done using 62.5 nM of miRCURY LNA miRNA inhibitor against has-miR-92b-5p (Qiagen, Germany) and miRCURY LNA miRNA Inhibitor Control (Qiagen, Germany) was used as the inhibitor control. Transfections were done using Opti-MEM media (Invitrogen, USA) for 5 h and replaced with complete MEM. Cells were processed after 48 hours of transfection for RNA and Protein analysis. Effective doses of mimic and inhibitor miRNA were standardized via qPCR and protein studies.
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