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. Author manuscript; available in PMC: 2020 Dec 11.
Published in final edited form as: Hepatology. 2020 Aug 26;72(4):1412–1429. doi: 10.1002/hep.31414

Figure 2: Telomere shortening impairs hepatocyte development in DKC1_A353V hESCS.

Figure 2:

(A, B) Endoderm generation from WT and DKC1_A353V hESCs at early (EP) and late (LP) passages, as assessed by (A) formation of CXCR4+CD117+ population by flow cytometry (% of population of interest indicated in red, top right) and (B) relative expression (analyzed by real-time quantitative PCR) of the endoderm markers FOXA2 and SOX17. (C) Generation of a hepatic endoderm population quantified by the relative gene expression of HNF4α (by real-time quantitative PCR) in WT and DKC1_A353V cells at different passages. (D) Relative expression of hepatocyte markers (by real-time quantitative PCR) after 21 days of differentiation in WT and DKC1_A353V cells at different passages. (E) Immunocytochemistry showing expression of different markers (indicated in the figure) during differentiation of WT and DKC1_A353V cells in early or late passages. The specific day during differentiation is indicated on the right. Scale Bars represent 50 μM. (F) Quantification of albumin positive cells by immunofluorescence after 21 days of differentiation in WT and DKC1_A353V cells at different passages. A total of 1000 cells/slide was counted. (G) Quantification of albumin secretion (by ELISA) after 21 days of differentiation in WT and DKC1_A353V cells at different passages. n=3, mean ± SEM, *p≤0.05; **p≤0.0025; ****p≤0.0001. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test.