Fig. 3

Dlk1⁺ cells in HSDCs could lose Dlk1^M and transform into Dlk1⁻ cells. a–d Dlk1⁺ and Dlk1⁻ cells were sorted from E12.5 HSDCs via FACS and then cultured on type I collagen-coated dishes. After three passages, the cells were collected, and the expression of Dlk1 was detected by FACS (a). Dlk1⁺ cells and Dlk1⁻ cells derived from Dlk1⁺ cells were sorted and collected via FACS, and then, RT-PCR (b) and quantitative real-time PCR (c) were performed to measure the mRNA expression level of Dlk1, and western blotting (d) was used to detect the protein expression level of Dlk1 isoforms. HSDCs isolated from E12.5, E14.5, and E16.5 mouse fetal livers were collected, and the expression level of Dlk1 was determined by quantitative real-time PCR (e) and western blotting (f). Representative images from one of three experiments are shown, and the data are shown as the mean ± SEM of three independent experiments. **P < 0.01 (Mann-Whitney test in c and one-way ANOVA with post Dunn’s multiple comparisons test in e)