Fig 3. Hematopoietic-restricted loss of Zeb1 decreases hematopoietic colony formation potential and ability to compete with wt HSPCs for contribution to all hematopoietic lineages.

(A) HSPCs isolated from Vav-iCre; Zeb1^fl/fl fetal livers show decreased numbers of colonies in methylcellulose-based colony assays (first) that further decreases upon secondary replating (second) compared to Cre negative controls. (B) Schematic of competitive BM reconstitution experiments whereby equal numbers of BM cells from Zeb1 null CD45.2⁺ mice were mixed with equal numbers of control CD45.1⁺ cells and used to reconstitute lethally irradiated CD45.1⁺ mice. If Zeb1 null CD45.2+ HSPCs are not compromised, they would be expected to contribute equally as the control CD45.1+ cells in their contribution to all hematopoietic cells (equal CD45.2⁺-orange/CD45.1⁺-blue, top row) whereas if they are severely compromised then the control CD45.1⁺ cells will solely contribute to the reconstituted hematopoietic system (all CD45.1⁺- blue, lower row). (C) Zeb1 null (Vav-iCre, Zeb1^fl/fl) CD45.2⁺ donor cells (orange bars) were outcompeted by control CD45.1⁺ competitor HSPCs (blue bars) for their ability to contribute to all hematopoietic cells analyzed in the PB and BM (rightmost panels). Zeb1 ^fl/+ heterozygous (middle panels) and Cre negative (left panels) CD45.2⁺ doner cells in general contributed equally as well as the competitor CD45.1⁺ cells for their contribution to the hematopoietic system of recipient mice with the exception to the T-cell lineage. Data are represented as mean + SD from 4 biological replicates. *p < 0.05; **p < 0.01, nonparametric t test. Raw data for (A) are included in S1 Data. Raw data for (C) are included in B of S1 Table. BM, bone marrow; HSPC, hematopoietic stem and progenitor cell; PB, peripheral blood; wt, wild-type.