Figure 6.

Lysine deprivation or treatment with cladosporin affects the growth of patient‐devied CC organoids. (A) Relative expression of KARS1 mRNA in patient CC organoids and human healthy liver organoids (HLO) (n = 5‐11). Effects of (B) lysine deprivation or (C) cladosporin treatment on cell viability were measured by Alamar blue assay after culturing 3 or 7 days (n = 4‐12). Organoids were treated with (D) lysine deprivation or (E) cladosporin (100 µM) for 7 or 14 days. The results depicts the mean viability of CC organoids (CCO‐1, CCO‐2, CCO‐3) compared with the mean viability of healthy liver organoids (HLO‐1, HLO‐2, HLO‐3) (n = 18‐24). (F) Lysine deprivation or (G) cladosporin treatment affects organoid initiation as assayed 2 weeks following seeding. The number (n = 4‐10) and size (n = 18‐45) of organoids after culturing for 14 days were calculated. Quantification of the data target groups were relative to the negative control group. Data are presented as mean ± SEM and dots represent individual organoid cultures. *P < .05, **P < .01, ***P < .001, by the Mann‐Whitney test. CCO, cholangiocarcinoma organoid; HLO, human liver healthy organoid; FC, fold change