Genome-Wide Mapping of Bivalent Histone Modifications in Hepatic Stem/Progenitor Cells

Figure 4

Loss-of-function assays of Sox4 in Dlk⁺ cells. (a) Real-time RT-PCR analysis of Sox4 in the fetal liver (ED 11.5, ED14.5, and ED17.5), neonatal liver, and adult liver. Statistically significant (). (b) Western blot analyses in Sox4-knockdown cells using anti-Sox4 and anti-tubulin (loading control) antibodies. (c) The number of large colonies containing >100 cells at day 5. Statistically significant (). (d) The percentages of Alb⁺CK7⁺ bipotent cells in large colonies at day 5 of culture are shown as mean values for 10 colonies. Statistically significant (). (e) Fluorescence micrographs of large colonies (containing >100 cells) transduced with indicated viruses at day 5 of culture. Dual immunostaining was performed to detect Alb (red) and CK7 (green) expression. Nuclear DAPI staining (blue) is shown in the insets. Scale . (f) Immunostaining analyses demonstrated Casp3 (red) and nuclear DAPI (blue). Scale . (g) Real-time RT-PCR analysis of Sox9 in Dlk⁺ cell-derived colonies. Statistically significant (). (h) Bright-field images and fluorescence micrographs (inset panels) of cells in EHS gel culture for hepatocytic differentiation and collagen gel culture for cholangiocytic differentiation. Scale .

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