To evaluate the function of JMJD2C on MALAT1 promoter activity, A549 cells were transfected with sh-NC or sh-JMJD2C with a luciferase reporter plasmid containing MALAT1 promoter through Lipofectamine 3000 (Invitrogen). After 48 h, cell lysates were subjected to analysis of relative luciferase activity on the dual luciferase reporter gene detection system (Promega, WI, USA) [10 (link)].
The interaction between MALAT1 and miR-503-5p and the targeting relationship between miR-503-5p and SEPT2 were conformed. MALAT1 wild-type (WT), MALAT1 mutant (MUT), SEPT2 WT and SEPT2 MUT potentially binding to miR-503-5p were inserted into pGL4 luciferase reporter and co-transfected with miR-503-5p mimic or mimic-NC into A549 cells. After 48 h, the relative luciferase activity was measured using a dual luciferase detection system (Promega).
The interaction between MALAT1 and miR-503-5p and the targeting relationship between miR-503-5p and SEPT2 were conformed. MALAT1 wild-type (WT), MALAT1 mutant (MUT), SEPT2 WT and SEPT2 MUT potentially binding to miR-503-5p were inserted into pGL4 luciferase reporter and co-transfected with miR-503-5p mimic or mimic-NC into A549 cells. After 48 h, the relative luciferase activity was measured using a dual luciferase detection system (Promega).
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