Figure 3: MC-RR induces progressive and irreversible lumen collapse.

A) MC-RR (400 nm) induced increasing neonatal EHBD cholangiocyte spheroid collapse with increasing treatment time. Cell-cell contacts, as marked by ZO-1 staining, were maintained (white arrowhead) as the damage progressed. B) Quantification showing progressive lumen collapse with increasing MC-RR exposure time. C) Quantification showing that MC-RR-induced damage was irreversible after 15 h or 24 h exposure and that lumen patency did not improve even after MC-RR washout. All images are representative of at least 3 independent experiments, with at least 18 spheroids quantified for lumen closure. Control spheroids are vehicle treated. Red: actin, Green: ZO-1, Blue: DAPI. All scale bars 50 μm.