Potential binding sites of miR-194-5p and circ_0006168 or JMJD1C were provided by Circinteractome (https://circinteractome.nia.nih.gov/) and TargetScan (http://www.targetscan.org/). The fragments of circ_0006168 and JMJD1C 3’UTR that contained miR-194-5p binding sites were synthesized and inserted into pmirGlO luciferase reporter vector (Promega, Madison, WI, USA), namely wild-type reporter vectors (circ_0006168-wt and JMJD1C-wt). Meanwhile, their mutated-type reporter vectors (circ_0006168-mut and JMJD1C-mut) without binding sites were constructed in the same way. Each of the above-mentioned vectors was respectively transfected into Eca109/Taxol and KYSE150/Taxol cells along with miR-194-5p or miR-NC for 48 h. The luciferase activity was estimated through a Dual-luciferase Reporter Assay System (Promega).
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