EKLF-null primitive erythroid cells show defects in gene expression and structure. (A) Relative expression levels of putative EKLF target genes in E11.5 EKLF^-/- blood cells compared with EKLF^+/+. Levels were measured by real-time PCR and normalized to HPRT. Data are presented as mean, n = 2. (B) Representative Western blot of dematin and AHSP expression in whole-cell extracts from E11.5 blood cells. Actin is presented as a loading control. (C) Representative dot plots of FACS-sorted E11.5 blood cells from EKLF^+/+, EKLF^+/-, and EKLF^-/- mice stained with TER119-PE and either CD71-FITC (transferrin receptor) or CD117-FITC (c-kit). The top and right quadrants represent positive stained cells. The percentage of cells in each quadrant is indicated. (D) Scanning electron microscopy of cytospin preparations of primitive erythroid cells from E11.5 EKLF^+/+ and EKLF^-/- embryos. The small enucleated discs represent maternal red cells and the large nucleated cells are yolk sac-derived primitive erythrocytes. Scale bars and magnification are shown.