Visualization of functional bile canaliculi
All organoids utilized for bile canaliculi visualization were subjected to standard preparation consisting of a single wash with DPBS containing calcium and magnesium (DPBS Ca/Mg) (Gibco) followed by Hoescht staining in Clonetics™ HCM™ Hepatocyte Culture Medium (HCM) (Lonza) for 30 mins at 37°C. For snapshot imaging of the bile canaliculi, organoids were incubated in HCM media containing 1uM of CDFDA (Molecular Probes) for 2h at 37°C, washed twice with DPBS Ca/Mg and transferred into a well of a µ-slide Angiogenesis glass bottom plate (Ibidi) containing HCM. For time-lapse imaging, organoids were kept constantly in HCM media containing 1uM of CDFDA (HCM-CDFDA). Organoids were incubated with indicated concentrations of troglitazone (Sigma Aldrich) at both 37°C and room temperature as specified.