Glut1 expression during erythroblast maturation is age-dependent. (A) Freshly isolated splenocytes obtained from 2-day-old, 14-day-old, and adult mice were stained with CD71 and Ter119 antibodies; density plots are shown. The percentages of cells in each region indicated in each plot, broadly correspond to proerythroblasts (region I), basophilic erythroblasts (region II), late basophilic/polychromatophilic erythroblasts (region III), and orthochromatophilic erythroblasts (region IV), as previously described.22 The relative expression of Glut1 in each of the 4 regions was determined by gating on the identified CD71/Ter119 populations. Shaded histograms show nonspecific staining. (B) At 2 days of age, splenic cells from regions I to IV were FACS-sorted as indicated in the dot plots in panel A. Total levels of Glut1 in each population were assessed by immunoblotting with a polyclonal anti-Glut1 antibody. Protein levels in each lane were controlled by staining with an antiactin antibody. (C) Erythroblast maturation in the bone marrows of 5-day-old, 14-day-old, and adult mice was assessed as described. The relative percentages of erythroid precursors in regions I to IV are indicated in dot plots showing CD71 staining as a function of Ter119. At each age, histograms showing surface Glut1 staining in the 4 regions are presented.
