Transient transfection and luciferase reporter assays

RAW264.7 cells were transiently co-transfected with KLF9-Luc or COX-2-Luc and the respective expressed plasmid (GR or KLF9) using Lipofectamine 2000 (product no. 11668019; Invitrogen; Thermo Fisher Scientific, Inc.). After 48 h, the cells were treated with Dex or LPS for another 3 h. Then, reporter gene assays were performed using a luciferase assay system (product no. E2810; Promega Corp.). The internal reference was determined by measuring the TK (mammalian vector for weak constitutive expression of humanized Renilla luciferase) bioluminescence intensity.