P450 glo cyp1a2 induction inhibition assay

In the ICG uptake assay, bioprinted structures or cells were cultured for 48 hours in a medium supplemented with 8-bromo-cAMP, pregnenolone-16α-carbonitrile, and progesterone, as per the manufacturer's instructions. Subsequently, they were coincubated with ICG (1 mg/ml) at 37°C for 1 hour, washed three times with PBS, and then switched to HEM culture medium. The uptake/release of ICG was observed under a microscope. Bioprinted structures or cells were stained with ac-LDL (Yeasen) and periodic acid-Schiff (Solarbio), and the staining results were observed under a microscope. Different inducers were used to treat 3DP-liver and eHep cells for 48 hours, and the gene expression levels of Cyp1a2 (3-methylcholanthracene, 10 μM, Sigma-Aldrich), Cyp3a11 (rifampicin, 25 μM, Solarbio), Cyp2e1 (rifampicin, 25 μM, Solarbio and ethyl alcohol; 0.5%, v/v), Cyp3a41b (3-methylcholanthracene, 10 μM, Sigma-Aldrich), and Cyp1a1 (rifampicin, 25 μM, Solarbio and barbituric acid, 10 μM, Aladdin) were detected. The P450-Glo Cyp1a2 Induction/Inhibition Assay (Promega) was used, following the manufacturer's instructions, to detect Cyp1a2 enzyme activity in different states.