IO induces inflammatory responses in hepatocyte cell lines in vitro. (A) Cells were treated with 30 μM apo-Tf or holo-Tf. mRNA expression levels were determined 24 hours posttreatment. (B) Various human hepatic cell lines and Thp-1–derived macrophages were treated with 30 μM apo-Tf or holo-Tf. Total mRNA was isolated 24 hours posttreatment. (C-D) Huh7 (C) and HL-7702 (D) cells were either mock treated (Cont.) or treated with 30 μM apo-Tf or holo-Tf. Protein levels of various inflammatory factors were determined 24 hours posttreatment. (E-F) Human primary hepatocytes were treated with holo-Tf at the indicated concentrations and apo-Tf as an appropriate control. Total RNA and proteins were isolated 24 hours posttreatment. Data were normalized to the control group and are presented as mean ± standard error of the mean (SEM) (N = 4). *P < .05; **P < .01 vs control.