In addition, we categorized drugs where no hepatic ADRs appeared sellekchem in any of the ADR sections as ��not mentioned�� (NM). To verify that the classifications from the automated search were correct, a low threshold search using ��liver�� and ��hepatic�� as search keywords followed by manual inspection of matching drug labels was conducted. The ADR sections in FDA drug labels describe ADRs in a hierarchical way on the basis of their severity. To assure that the ADR sections described DILI severity in the same hierarchical way, without influence from other ADRs reported for the same drug, the severity of DILI reported in the different sections was evaluated using the approach described in Chen et al. (2011) and Supplementary Table S2.

Because FDA drug labels of registered drugs were used to investigate the correlation between BSEP inhibition and DILI, the impact on the data set composition from the exclusion of nondrug compounds (n = 68) was assessed. The frequency of BSEP inhibitors was comparable in the full data set (n = 250) investigated for BSEP inhibition in membrane vesicles (34% BSEP inhibitors) and in the subset of drugs (n = 182) in the DILI analysis (33% BSEP inhibitors). Furthermore, the exclusion of nondrugs did not skew the data set with regard to chemical diversity, as confirmed by a principal component analysis of molecular descriptors (data not shown). Isolation and culture of human hepatocytes. Liver tissues free from metastases were obtained from human donors undergoing partial liver resections at the Department of Surgery, Uppsala University Hospital, Sweden (see Table 1 for patient demographics).

All donors gave informed consent, in accordance with the approval from the Uppsala Regional Ethical Review Board (Ethical Approval no. 2009/028). Hepatocytes were isolated using the 2-step liver digestion technique of Lecluyse and Alexandre (2010). Primary hepatocytes, with a viability >85%, were seeded in collagen-1-coated 24-well plates (BD Biosciences) at a density of 3.75��105 cells per well and initially maintained at 37��C and 5% CO2 in 500 ��l DMEM supplemented with 5% (vol/vol) fetal bovine serum, 4 ��g/ml insulin, 1��M dexamethasone, 4mM l-glutamine, 100U/ml penicillin, and 100 ��g/ml streptomycin. Cells were allowed to attach to the plate for 2�C3h in a humidified culture chamber at 37��C and 5% CO2, after which the medium was carefully aspirated and replaced with 500 ��l culture medium (HMM supplemented with insulin 10 ��g/ml, transferrin 5.

5 ��g/ml, selenium 5ng/ml, 0.1��M dexamethasone, 100U/ml penicillin, and 100 ��g/ml streptomycin). After overnight incubation, the cells were overlaid with 500 ��l ice-cold 0.25mg/ml Matrigel in culture medium. To allow the formation of bile canaliculi, the cells were cultured for an additional GSK-3 4�C6 days during which the culture medium was refreshed every 24h.