For miRNA transfection, 293T cells or OPCs were incubated with 50 nM miR-344e/410-3p mimic/agomir or antagomir using Lipofectamine 3000 Reagent (Thermo Fisher Scientific). An equal concentration of a nontargeting control sequence was added to experimental samples as controls for non–sequence-specific effects in miRNA experiments.

The entire and mutated 3′ UTR of human and mouse LDLR were generated by Ribo Bio. 293T cells were plated into 96-well plates and cotransfected with 1 μg of the indicated 3′ UTR luciferase reporter vectors and 100 nM miR-344e/410-3p mimics or control mimic (Ribo Bio). Renilla and firefly luciferase activities were measured using the Dual-Luciferase Reporter Assay System (E1910, Promega) with the GloMax-Multi+ Detection System (Promega). Experiments were performed in duplicate wells and repeated 4 times.

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