ZNF652-Induced circRHOT1 Promotes SMAD5 Expression to Modulate Tumorigenic Properties and Nature Killer Cell-Mediated Toxicity in Bladder Cancer via Targeting miR-3666

Figure 6

miR-3666 directly targets SMAD5. (a) RNA pull-down assay conducted using miR-3666 probe, followed by qRT-PCR, to measure miR-3666 expression. (b, c) Putative mRNA expression in HT-1376 (b) and HTB9 (c) cells determined by RNA pull-down assay using miR-3666 probe and qRT-PCR. (d, e) Expression of SMAD5 in HTB9 (d) and HT-1376 (e) cells measured by RNA pull-down assay using biotin-miR-3666-WT/Mut probe combined with qRT-PCR. (f) The binding sites between SMAD5 and miR-3666. (g, h) Relative luciferase activities in HT-1376 (g) and HTB9 (h) cells cotransfected with miR-3666 mimics/NC mimics and WT/Mut SMAD5 plasmids. (i) Relative mRNA expression of SMAD5 in HT-1376 or HTB9 cells transfected with miR-3666 mimics or inhibitors measured by qRT-PCR. (j) Expression of SMAD5 protein in HT-1376 or HTB9 cells transfected with miR-3666 mimics or inhibitors detected by western blot. Experiments were conducted in triplicate. and .

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