The target miRNAs for PLOD1 were selected by TargetScan (http://www.targetscan.org/vert_72/ ), based on the context++ score (22 (link)). The plasmids of the wildtype p3'-UTR-PLOD1 and the mutant p3'-UTR-PLOD1-mut were generated. For the dual-luciferase reporter assay, plasmids were co-transfected into AMSCs using LipofectamineTM 2000 system (Invitrogen, USA).
Protocol detail
Identifying PLOD1 miRNA Targets
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Corresponding Organization : Shanghai Ninth People's Hospital
Other organizations : XinHua Hospital, Changhai Hospital, Second Military Medical University
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Variable analysis
independent variables
- Wildtype p3'-UTR-PLOD1 plasmid
- Mutant p3'-UTR-PLOD1-mut plasmid
- Luciferase reporter activity
- Cell type (AMSCs)
- Transfection method (Lipofectamine 2000)
- Positive control: Not specified
- Negative control: Not specified
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