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. 2020 Apr 24;11(4):278. doi: 10.1038/s41419-020-2485-1

Fig. 5. KCNQ1OT1 bound to miR-34c-5p and decreased expression in the OS cells.

Fig. 5

a The expression of miR-34c-5p was upregulated in U-2OS and143B transfected with KCNQ1OT1 shRNA or the control shRNA by RT-qPCR, **p < 0.01, ***p < 0.001 (Student’s t-test). b The wild-type and the mutated sequences of the KCNQ1OT1 mRNA 3’-UTR (mutation site: red). c Top panel shows a schematic image of a construction containing KCNQ1OT1 wild type combined with MS2 binding sequence. MS2-RIP followed by miR-34c-5p qRT-PCR to measure miR-34c-5p endogenously associated with KCNQ1OT1, **p < 0.01 (Student’s t-test). d U-2OS and 143B cells lysate were incubated with biotin-labeled KCNQ1OT1, qRT-PCR measured miR-34c-5p expression in the products of pulldown by biotin, **p < 0.01 (Student’s t-test). e and f The luciferase activity of the OS cells (U-2OS and 143B) in luciferase reporter plasmid containing wild-type KCNQ1OT1 3’-UTR (KCNQ1OT1-WT) and mutant KCNQ1OT1 3’-UTR (KCNQ1OT1-MUT) co-transfected with miR-34c-5p mimics or negative control was assessed, ***p < 0.001 (Student’s t-test). g and h AGO2-RIP followed by qPCR to evaluate KCNQ1OT1 level after miR-34c-5p overexpression, **p < 0.01, ***p < 0.001 (Student’s t-test).