Time and dose-dependent changes in HepaRG cells with APAP treatment for ALT, APAP protein adducts, miRNA (miR-122-5p, miR-378a-5p, miR-27b-3p, and miR-125b-5p and mRNA levels (CYP2E1, CYP3A4, and CYP1A2) levels. Effect of 5 and 20 mM APAP treatment of HepaRG cells at 1, 6, 12 and 24 h time points. (A and B) Show levels of ALT and APAP protein adducts in cell medium; *p < 0.05 compared to controls for APAP 5 mM and 10 mM. Parenthesis around the asterisk denotes significance of ALT for 5 mM APAP treatment at 12 and 24 h. (C and D) Depict miRNA expression in cell medium. Data were normalized for HepaRG cell culture medium with Let-7d and spiked-in C. elegans miR-39; *p < 0.05 compared to controls. (E and F) Describe CYP1E2, CYP1A2 and CYP3A4 mRNA levels determined by qRT-PCR. TaqMan assays were duplexed with GAPDH to normalize the mRNA expression. Parenthesis around the asterisk differentiate 24 h CYP3A4 from CYP1A2; *p < 0.05 compared to controls. “h” Denotes hour. Error bars represent Standard error of the mean.