Figure 3.
Figure 3. Tie2-Cre mediated deletion of Ezh2 leads to a specific loss of mKitL expression. (A) KitL and Epo mRNA expression levels in E12.5 FL cells (WT, n = 6; KO, n = 4; 2 independent experiments). (B) Gating strategy used to define hepatoblast (Ter119−CD45−CD31−Dlk-1+) population within the total FL live (7AAD−) singlet population. (C) Proportion of hepatoblasts in E12.5 FL (WT, n = 10; KO, n = 11; 4 independent experiments). Two-tailed Student t tests were used to assess statistical significance. Error bars represent SEM. (D-E) Immunofluorescence staining of E12.5 FL (n = 3 each) using antibodies against KitL (D, red), Alb (D, green, hepatoblast marker), c-Kit (E, magenta, hematopoietic cell marker), CD31 (E, red, endothelial cell marker), mKitL (E, green). Nuclei was stained with 4′,6-diamidino-2-phenylindole (DAPI; D-E, blue). Scale bars, 20 µm. (F) Representative FACS histogram plots showing the recombination efficiency of Tie2-Cre in E12.5 FL hepatoblasts (n = 5, 2 independent experiments) and endothelial cells (n = 6, 1 experiment; Ter119−CD45−CD31+Lyve1+) using Rosa26-LSL-tdTomato reporter. Shown are the mean values with SEM for the frequencies of the indicated population across all experiments.

Tie2-Cre mediated deletion of Ezh2 leads to a specific loss of mKitL expression. (A) KitL and Epo mRNA expression levels in E12.5 FL cells (WT, n = 6; KO, n = 4; 2 independent experiments). (B) Gating strategy used to define hepatoblast (Ter119−CD45−CD31−Dlk-1+) population within the total FL live (7AAD−) singlet population. (C) Proportion of hepatoblasts in E12.5 FL (WT, n = 10; KO, n = 11; 4 independent experiments). Two-tailed Student t tests were used to assess statistical significance. Error bars represent SEM. (D-E) Immunofluorescence staining of E12.5 FL (n = 3 each) using antibodies against KitL (D, red), Alb (D, green, hepatoblast marker), c-Kit (E, magenta, hematopoietic cell marker), CD31 (E, red, endothelial cell marker), mKitL (E, green). Nuclei was stained with 4′,6-diamidino-2-phenylindole (DAPI; D-E, blue). Scale bars, 20 µm. (F) Representative FACS histogram plots showing the recombination efficiency of Tie2-Cre in E12.5 FL hepatoblasts (n = 5, 2 independent experiments) and endothelial cells (n = 6, 1 experiment; Ter119−CD45−CD31+Lyve1+) using Rosa26-LSL-tdTomato reporter. Shown are the mean values with SEM for the frequencies of the indicated population across all experiments.

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