Biotin-labeled circMID1 probe, the wild-type (wt)/mutate-type (mut) of miR-330-3p probe (bio-miR-330-3p-wt/mut) and their negative control probes (NC probe and bio-NC) were synthesized by Genepharma. PC-3 and DU145 cells were harvested, and then the cell lysates were incubated with Dynabeads M-280 Streptavidin (Invitrogen) coated circMID1 probe or NC probe. The enrichment of candidate miRNAs was examined by qRT-PCR to screen the targeted miRNA for circMID1. To explore whether miR-330-3p could pull down circMID1, YTHDC2 and IGF1R, PCa cells were transfected with the bio-miR-330-3p-wt/mut probe or bio-NC probe for 24 h, as the previous study [16 (link)]. Then, the cell lysates were cultured with the Dynabeads, and the RNA enrichment was analyzed by qRT-PCR.
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Identifying circMID1-Targeting miRNAs in Prostate Cancer
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Corresponding Organization : First Affiliated Hospital of Zhengzhou University
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Variable analysis
independent variables
- Transfection of PC-3 and DU145 cells with bio-miR-330-3p-wt/mut probe or bio-NC probe
- Enrichment of candidate miRNAs examined by qRT-PCR
- Enrichment of circMID1, YTHDC2, and IGF1R analyzed by qRT-PCR
- Incubation of cell lysates with Dynabeads M-280 Streptavidin coated with circMID1 probe or NC probe
- Biotin-labeled circMID1 probe
- Bio-miR-330-3p-wt probe
- NC probe
- Bio-NC probe
- Bio-miR-330-3p-mut probe
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