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. 2014 Aug 25;9(8):e104776. doi: 10.1371/journal.pone.0104776

Figure 1. Ezh2 is highly expressed in parenchymal epithelial cells in early developing liver.

Figure 1

A: Time course analysis of expression levels of the polycomb group (PcG) members (Ezh1, Eed, Suz12, Bmi1, Ring1B, and Ezh2) in CD45− TER119− non-hematopoietic liver cells of mice at nine time points between ED 9.5 and 8 weeks. Relative expression values from microarray analyses are shown. B: Western blot analysis of Ezh2 in CD45− TER119− non-hematopoietic liver cells of fetal mice at ED 11.5, 13.5, 15.5, and 17.5. β-actin is indicated as control. C: Immunofluorescence staining for Ezh2, CK8/18 (epithelial marker), and DAPI in liver of fetuses at ED 11.5, 13.5, 15.5, 17.5, P 0 (neonatal), and 8 weeks. Arrows indicate Ezh2 positive cells in CK8/18 positive cells. (Scale bar: 100 µm) D: Frequency of Ezh2 positive cells in CK8/18 positive cells at indicated time points. (n = 3). E: Immunofluorescence staining for histone H3K27me3, CK8/18 (epithelial marker), DAPI in the liver tissues at ED 11.5, 13.5, 15.5, and 17.5. Scale bar = 100 µm. F: Frequency of H3K27me3 expressing cells in CK8/18 positive cells at indicated points. Data are mean ± SD (n = 3). G Immunofluorescence staining for Ezh2, H3K27me3, Ck8/18 and DAPI in the liver of fetuses at ED 13.5. Upper is merge image of all colors. Bottom column shows image of Ezh2 and H3K27me3, lower right is magnified image of left. Red: Ezh2, Green: H3K27me3, Light blue: CK8/18, Blue: DAPI. Scale bar = 100 µm. H Immunofluorescence staining for AFP, Ezh2, CK8/18, and DAPI in the liver of fetuses at ED 13.5. Arrows indicate AFP and Ezh2 double positive cells in CK8/18 positive cells. (Scale bar: 50 µm) I: Frequency of Ezh2 positive cells in AFP and CK8/18 double positive cells and AFP negative and CK8/18 positive cells at indicated time points. (n = 3). P values (asterisks) are from the Mann–Whitney U-test. *P<0.05.