Firefly Luciferase Reporters for Il9 and Irf4

Reporter vector coding for the Firefly Luciferase under the control of the Il9 promoter encompassing nucleotides −1201 to +52 bp was cloned into the promoterless pGL3 Basic luciferase reporter gene vector (Promega)^{28 (link)}. Irf4 promoter encompassing nucleotides −1562 to +122 bp was cloned into pGL3 Basic luciferase reporter gene vector. Reporter assays were carried out as described previously^{7 (link)}. Briefly, 293 T cells were transfected with 0.4 μg of the reporter vector coding for the Firefly Luciferase under the control of the Il9 or Irf4 promoter and with 0.8 μg of the Foxo1 vector. Cells were cultured for 48 hours before harvesting and the relative Il9 or Irf4 promoter activity was measured using Promega kit in accordance with the manufacturer’s instructions.