Figure 6: MC-RR induces elevated reactive oxygen species in neonatal EHBD cholangiocyte spheroids.

A) Quantification of Ki67+ cells in neonatal mouse EHBD cholangiocyte spheroids treated with vehicle or 400 nM MC-RR (n=18 per condition from 3 independent experiments). B) Spheroid diameter after treatment with vehicle or 400 nM MC-RR (n=45 spheroids per condition from 3 independent experiments). C) Relative retention of MC-LR and MC-RR in neonatal EHBD cholangiocytes (n=9 samples from 3 independent experiments). D) MC-RR retention in neonatal and adult EHBD cholangiocytes (n=9 samples from each condition across 3 independent experiments). E) ROS production in neonatal and adult EHBD cholangiocyte spheroids after treatment with 400 nM MC-LR or 400 nM MC-RR for 24 h. F) ROS production in the presence of vehicle or 400 nM MC-RR. Images are representative of 3 independent experiments with at least 18 spheroids quantified for lumen closure per condition. Red: actin, Green: Cell-Rox Green, Blue: DAPI. All scale bars 50 μm.