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. 2019 Jan 22;2019:5968236. doi: 10.1155/2019/5968236

Figure 5.

Figure 5

Hepatocyte differentiation of 3 μM CHIR-derived DE cells (6 d). (a) DE cells derived from H9 cells treated with 3 μM CHIR for 48 hr in DE media followed by 4 days in CHIR-withdrawn DE media (3 μM CHIR (6 d)) were cultured in hepatic progenitor media without 100 ng/ml HGF added for 7 days and then cultured in Touboul's maturation media for another 7 days. The cells were fixed on day 13 and day 20 of differentiation and photographed for phase images. The cells were then stained and imaged by a fluorescence microscope using antibodies against AFP, HNF4α, and ALB. DAPI represents nuclear staining. Scale bar = 100 μm. (b) The cells as explained in (a) were analyzed for mRNA expression by RT-qPCR for the indicated genes using gene-specific primers. The bars represent normalized (18S rRNA) fold mRNA expression. The data are represented as mean ± standard deviation.