Primary myoblasts were transfected 48 h with 10 nM of miRIDIAN miRNA mimics for mmu-miR-208b-3p or mmu-miR-499-5p or negative control (miR-Ctrl) (Dharmacon) using Dharmafect 3 reagent (Dharmacon). For transient transfections, 10T1/2 cells were seeded 24 h on 24 well plates before transfections at 50 to 60% confluence. For luciferase reporter assays, 10T1/2 cells were co-transfected with 4 nM of miRNA mimics or negative control (miR-Ctrl) (Qiagen), 4 nM miScript col25a1 target protector or negative control (Qiagen) and 0.1 µg/well of luc-Col25a1 or mutated luc-Col25a1 plasmids using Dharmafect Duo. 0.1 µg/well of pCMV-Renilla plasmid were included as an internal control in all transfection assays. Cells were harvested and lysed 48 h after transfection and luciferase activity was analyzed using the Dual Luciferase Reporter Assay System (Promega).
Mir ctrl
Manufactured by Horizon Discovery
The MiR-Ctrl is a laboratory equipment product designed for the manipulation and control of microRNAs (miRNAs). It provides a platform for researchers to study the function and regulation of miRNAs, which are small, non-coding RNA molecules that play a crucial role in gene expression and cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Dharmafect 3 reagent, by Horizon Discovery (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
A2780, by American Type Culture Collection (1 mentions)
Small interfering rna sirna, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Skov3, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Heya8, by American Type Culture Collection (1 mentions)
Scrambled negative sirna control, by Merck Group (1 mentions)
2 protocols using mir ctrl
1
Myoblast miRNA Transfection and Luciferase Assay
2
Ovarian and Cervical Cancer Cell Lines
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The ovarian cancer cell lines SKOV3, HeyA8 and A2780, and the cervical cancer cell line HeLa were obtained from the American Type Culture Collection and maintained in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) without any antibiotics. The cells were incubated at 37°C in an atmosphere containing 5% CO2. Human miR-508-3p mimic (hsa-miR-508-3p) and scrambled negative control (miR-ctrl) were obtained from GE Healthcare Dharmacon, Inc. Small interfering RNA (siRNA) targeting CCNA2 or MMP7 and scrambled negative siRNA control were purchased from Sigma-Aldrich; Merck KGaA. The pcDNA3.1 plasmids (Promega Corporation) expressing CCNA2 or MMP7 and empty vectors (EV) were constructed by Shanghai GenePharma Co., Ltd.
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