Fig. 2.

Genetic manipulation of AKR1D1 in human hepatoma cell lines and bile acid signalling and synthesis in human hepatoma cell lines, following AKR1D1 knockdown. AKR1D1 knockdown (white bars), decreased mRNA and protein expression in both HepG2 and Huh7 cells, as measured by qPCR and western blotting (a-c). In both HepG2 and Huh7 cells, AKR1D1 knockdown (white bars) decreased total bile acid concentrations (d). In addition, AKR1D1 knockdown decreased cholic acid (CA) and chenodeoxycholic acid (CDCA) concentrations in Huh7 cells (e). Consistent with these, AKR1D1 knockdown increased the mRNA expression of genes involved in classic bile acid synthesis pathway (CYP7A1, HSD3B7, CYP8B1), bile acid transport (SLC51A), and FXR activation (NR0B2, LRH-1) (f-g). Representative Western blot images are shown, however formal quantification was performed in n = 7–9 replicates. qPCR data were normalised to 18SrRNA. Data are presented as mean ± se of n = 5–8 experiments, performed in triplicate, *p < 0.05, **p < 0.01, ***p < 0.001, compared to scrambled controls (black bars).