Primary rat cardiomyocytes were provided by ATCC (Shanghai, China). To produce cells overexpressing the Smurf1 (the Smurf1 group), cells were delivered with the Smurf1 plasmid for 8 hours using 5 µL Lipofectamine 3000 (Invitrogen Co., LA, USA). Next, cells were cotransfected with Smurf1 plasmids and miR-129-5p mimic (30 µmoL; Riobio, Guangzhou, China) to produce the Smurf1 + miR-129-5p group. The proteasome inhibitor MG132 (10 µM) or DMSO (0.1%) was used as solvent control. They were added to the cell culture medium 2 hours prior to mechanical strain application and remained in the culture medium throughout the experiment [23 (link)].
Mir 129 5p
Manufactured by RiboBio
Sourced in China
MiR-129-5p is a microRNA (miRNA) product available at RiboBio. miRNAs are small, non-coding RNA molecules that play a regulatory role in gene expression. The core function of MiR-129-5p is to modulate the expression of target genes, though its specific applications are not provided here.
Automatically generated - may contain errors
Lab products found in correlation
Mir 129 5p mimic, by RiboBio (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Lipofectamine ltx reagent, by Thermo Fisher Scientific (1 mentions)
Pgl3 basic plasmid, by Addgene (1 mentions)
Prl tk renilla plasmid, by Promega (1 mentions)
Dual luciferase reporter assay kit, by Promega (1 mentions)
2 protocols using mir 129 5p
1
Overexpression of Smurf1 and miR-129-5p in Rat Cardiomyocytes
2
Validating miR-129-5p Regulation of OTX1
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MiR-129-5p or MiR-129-5p mut mimics were purchased form RiboBio Co., LTD (Guangzhou, China). DNA fragment form the 3′-untranslate region (UTR) of OTX1 containing the predicted complementary sites of MiR-129-5p were constructed into a pGL3-basic plasmid (Addgene, Cambridge, USA). 1 × 10 4 LSCC cells were seeded in 48-well plates in triplicate and settled for 24 h. Then, the pGL3-OTX1 -3'UTR reporter plasmids (100 ng) plus 5 ng of pRL-TK renilla plasmid (Promega, Madison, USA), and increasing amounts (10 and 50 nM) of negative control (NC), MiR-129-5p or MiR-129-5p mut mimic were cotransfected into LSCC cells using Lipofectamine LTX reagent (Invitrogen, Carlsbad, USA) according to the manufacturer's recommendation. Luciferase and renilla signals were measured 24 h after transfection using the Dual Luciferase Reporter Assay Kit (Promega, Madison, USA) according to a protocol provided by the manufacturer.
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