ARIEL recruits the mediator complex to the ARID5B −135-kb enhancer locus. (A-C) ChIP analysis was performed using a specific antibody against mediator 1 (A), mediator 12 (B), or RNA POL II (C) or control IgG in Jurkat cells after the shRNA knockdown of ARIEL. See Figure 1B legend for details. (D) Jurkat cells were subjected to a RIP assay using a specific antibody against mediator 1 or mediator 12 or control IgG. The RNA transcripts in the RIP lysate and input samples were measured by qRT-PCR using specific primers targeting ARIEL or GAPDH (negative control). The percentage of the input is shown as the mean plus or minus SD of triplicate experiments. (E) The sense or antisense strand of ARIEL was biotinylated and mixed with the whole-cell lysate from Jurkat cells. The in vitro pulldown assay was conducted and western blot analysis was performed to detect the presence of the mediator 12 protein in the pulldown lysate.