IO negatively regulates HNF4α and miR-122 expression in hepatocyte cell lines in vitro. (A) Huh7 and HL-7702 cells were treated as indicated. Total RNA was isolated 24 hours posttreatment. (B) Huh7 cells were treated with 30 μM apo-Tf or holo-Tf. Both groups were transfected with either miR-122 mimics or scramble controls. The protein levels of various inflammatory factors were determined 24 hours posttreatment. (C) Huh7 (left) and HL-7702 (right) cells were treated as indicated. Total RNA was isolated at 24 hours posttreatment. (D) Huh7 (top) and HL-7702 (bottom) cells were treated with 30 μM apo-Tf or holo-Tf. CCL2 and HNF4α protein expression was determined 24 hours posttreatment. (E) Huh7 cells were treated with 30 μM apo-Tf or holo-Tf. ChIP assays were performed using anti–HNF4α monoclonal antibody, followed by quantitative polymerase chain reaction analysis using primers specific to the miR-122 promoter region. Mouse immunoglobulin G (IgG) was used as a negative control. (F) Huh7 cells were treated with 30 μM apo-Tf or holo-Tf. Both groups were transfected with either HNF4α overexpression plasmids (oeHNF4α) or control plasmids. Total RNA was isolated 24 hours posttreatment. Data were normalized to the control group and are presented as mean ± SEM (N = 4). *P < .05; **P < .01 vs apo-Tf; #P < .05; ##P < .01 vs holo-Tf.