Fig 3. Enhancer activity of the 1 kb IL2RA intronic region containing rs12722489 depends on the single nucleotide variant and is estrogen-dependent.

(A) Design of the pGL3-based vectors used for luciferase reporter assay. The constructs contained either G or A variant of the 1kb fragment of IL2RA first intron downstream of the luciferase gene. The control vector (not shown on the figure) contained an irrelevant 1 kb sequence (see Materials and methods section for details). Position of rs12722489 within the 1kb intronic region is indicated. (B) MT-2 or Jurkat cells grown in complete RPMI medium were transiently transfected with the luciferase reporter constructs. *p<0.05 comparing to control or 1kb(A) construct. (C) MT-2 cells transfected with the constructs indicated in the legend were placed in steroid-free medium and 17β-estradiol (E2) was added 24h later at the specified concentrations. Luciferase signal was assessed 18h after E2 addition. *p<0.05 comparing to cells without E2. Data from at least 3 independent experiments are represented as mean±SEM.