Luciferase Reporter Assays for LXRα Transcriptional Activity Determination

Dual-luciferase reporter assay was used to evaluate the transcription regulatory activity of RELA on LXRα gene. Briefly, RELA response element (RELA RE) and either wild-type or mutated LXRα luciferase reporter vectors were transfected into the MKN45 cells, followed by the stimulation with 500 U/ml IL-2 alone or together with 10 μM JSH-23 for 24 h. In another experiment, the RELA response element (RELA RE) and either wild-type or mutated LXRα luciferase reporter vectors were transfected into MKN45 cells overexpressing XEDAR, and then the cells were treated with JSH-23. Finally, the luciferase activity was determined with a Promega kit (Promega, Madison, WI) and Multiskan FC Microplate Reader (Thermo Fisher Scientific, Waltham, MA, USA).