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. 2023 Jun 9;16(3):451–472. doi: 10.1016/j.jcmgh.2023.06.003

Figure 9.

Figure 9

AKT inhibition rescues DC phenotypes in hepatocytes.(A) Immunofluorescence images of HNF4α and Ki67 in HEPs treated with dibenzazepine (10 μmol/L), IWR-1-endo (10 μmol/L), CHIR99021 (3 μmol/L), 10058-F4 (50 μM), MK2206 (1.0 μmol/L), or dimethyl sulfoxide (DMSO) vehicle control starting at the definitive endoderm stage (day 5) and continuing until terminal HEP differentiation (day 17). Scale bar: 100 μm. (B) Lipid accumulation in HEPs analyzed by BODIPY staining. Scale bar: 100 μm. (C) Quantification of cell-cycle distribution in HEP cultures of indicated genotype and treatment. (D) Expression of hepatic marker genes (HNF4α, TDO2, ALB, and TTR), MYC, and TERT in 2D HEP cultures treated with AKT inhibitor MK2206 (n = 4). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001. Error bars indicate means ± SD. (E) Gene expression analysis of hepatic marker genes and MYC in 2D HEP cultures treated with MYC inhibitor 10058-F4 (n = 4). ∗P < .05. Error bars indicate means ± SD. (F) Immunofluorescence staining for HNF4α and Ki67 in 2D HEP cultures treated with indicated small molecules. Scale bar: 100 μm. (G) Western blot analysis of proteins involved in the AKT signaling pathway. (H) Schematic illustration of AKT signaling in hepatocytes. Cor, corrected; Mut, mutant.