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Fig. 4 | Cell & Bioscience

Fig. 4

From: Hsa-let-7c-5p augments enterovirus 71 replication through viral subversion of cell signaling in rhabdomyosarcoma cells

Fig. 4

Target of hsa-let-7c-5p in MAP4K4. a Predicted binding site of hsa-let-7c-5p to the 3′UTR of human MAP4K4, as determined using TargetScan. b MAP4K4 was validated as a target of hsa-let-7c-5p by dual luciferase reporter assay. RD cells were transfected with a wild-type or mutant 3′UTR vector (50 ng) for 6 h, followed by transfection of the NC (100 nM) or hsa-let-7c-5p mimic (100 nM) for 48 h. Renilla luciferase activity in each sample was normalized to Firefly luciferase activity. c The effect of the hsa-let-7c-5p mimic on the MAP4K4 mRNA level. RD cells were transfected with NC (100 nM) or hsa-let-7c-5p (100 nM) for 48 h. RNA was extracted and analyzed by qRT-PCR. d Regulation of the MAP4K4 protein level by the transfection of RD cells with the hsa-let-7c-5p mimic (100 nM) for 48 h, as demonstrated by Western blotting. The right panel in (d) shows that MAP4K4 expression normalized to GAPDH is expressed as a fold change compared with the NC mimic group, as quantified using Image J software. e The MAP4K4 mRNA level is affected by hsa-let-7c-5pi. RD cells were separately transfected with NCi (150 nM) and hsa-let-7c-5pi (150 nM) for 48 h, and then RNA was analyzed by qRT-PCR. f Regulation of the MAP4K4 protein level by the transfection of RD cells with hsa-let-7c-5pi (150 nM) for 48 h, as demonstrated by Western blotting. The right panel in (f) shows that MAP4K4 expression normalized to GAPDH is expressed as a fold change compared with the NCi group, as quantified using Image J software. *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance

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