Fig. 5

miR-524-5p regulates critical features of cellular reprogramming via targeting TP53INP1. a Endogenous transcript levels of miR-524-5p and TP53INP1 in WJ0706 cells. Before (mock) or after transfection of a miR-524-5p mimic, WJ0706 cells were harvested 48 h post-transfection for analysis; miR-524-5p copy number was determined by Taqman qRT-PCR (left panel); TP53INP1 transcript level was determined by direct RT-PCR (right panel). b–f The miR-524-5p mimic-transfected WJ0706 cells were subjected to assays to determine cell proliferation (b,c), cell viability (d), apoptosis after oxidative stress induced with 200 μM H₂O₂ (e), and expression of selected pluripotency genes (f). All the experiments also included mock transfection with a negative control (NC) miRNA mimic, or a TP53INP1 siRNA (siTP53INP1) included as controls. For effects on cell proliferation, cell counts at different days post-transfection (b), or by BrdU ELISA measurements (c) were performed. For apoptosis assay (d,e), cell viability 2 h after incubation with H₂O₂ was determined by the MTT assay (d), or the histone-associated DNA fragments of apoptotic cells were quantified by ELISA assay 6 h after H₂O₂ treatment (e). f Expression of pluripotency genes in the transfected cells was analyzed by real-time RT-PCR 48 h post-transfection. Relative absorbance unit and mRNA level were determined as the relative absorbance units and expression, respectively to the value of the mock and negative control experiments, respectively, which was set as 1.0. *p < 0.05, **p < 0.01