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. 2020 Jun 12;13:5495–5514. doi: 10.2147/OTT.S249168

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© 2020 Zhang et al.

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Dual-luciferase reporter gene assay. (A) Top: The 3ʹ-UTR sequence of SERTAD3 was fused to the luciferase gene. Indicated is the complementarity of the 3ʹ-UTR sequence with miR-92a. Also shown is the sequence of a mutant 3ʹ-UTR of SERTAD3 that has lost (indicated by red letters) its complementarity to miR-92a. Bottom: Relative luciferase assay comparing the PGL3-SERTAD3 and PGL3-SETRAD3-mut vectors in 293T cells co-transfected with miR-92a mimic, miR-92a inhibitor, and control (NC). Firefly luciferase activity was normalized to Renilla luciferase activity. **p < 0.01. Per condition, three independent experiments were performed. (B) Analysis of the 3ʹ-UTR sequences of the SERTAD family members identified SERTAD3 as the sole specific target of miR-92a. Green highlights in the SERTAD3 3ʹUTR sequence and in the miR-92a sequence indicate complementarity.