Validation of miR-19a-3p Targeting PHLDA3

The dual-luciferase enzyme reporter assay was conducted as previously documented [17 (link)]. The wild-type (WT) docking site of miR-19a-3p on PHLDA3 and the matching mutant (Mut) were cloned into the pGL3-basic vector. The PHLDA3 WT or PHLDA3 Mut was co-transfected into 143B/U2OS cells with miR-19a-3p mimics or inhibitor or their corresponding control. After 48 h, Promega Dual-Luciferases Reporter Assay kit (Promega E1980) was performed to measure luciferase activities according to the manufacturer’s protocols.

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Wang P., Huang Y., Xia X., Han J., Zhang L, & Zhao W. (2022). Pleckstrin homology-like domain family A, member 3, a miR-19a-3p-regulated gene, suppresses tumor growth in osteosarcoma by downregulating the Akt pathway. Bioengineered, 13(2), 3993-4009.

Publication 2022

Dual-Luciferases Reporter Assay kit

Assay Cells Enzyme assay Luciferase Pgl3 Promega Vector

Corresponding Organization : Second Affiliated Hospital of Dalian Medical University

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Variable analysis

independent variables

MiR-19a-3p mimics
MiR-19a-3p inhibitor

dependent variables

Luciferase activities

control variables

PHLDA3 wild-type (WT) docking site
PHLDA3 mutant (Mut) docking site
Promega Dual-Luciferases Reporter Assay kit
143B/U2OS cells

controls

PHLDA3 WT and PHLDA3 Mut co-transfected with corresponding control for miR-19a-3p mimics and inhibitor

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