Primary neonatal mouse cholangiocytes in spheroid culture were treated with vehicle (DMSO) or biliatresone for 24 h and immunostained for F-actin (green), the β1 integrin subunit (red), or ZO-1 (green) as indicated. b) A cholangiocyte cell line was treated similarly and immunostained for E-cadherin (green). c) Cholangiocyte cell line spheroids were treated with biliatresone for 1–12 h and immunostained for cellular tubulin (red) or ZO-1 (green). In all cases nuclei were stained with DAPI (blue). Scale bars, 10 μm. Representative images from 3 independent experiments, each experiment carried out in duplicate. BD – bile duct, PT – portal tract, EHBD – extra-hepatic bile duct