Figure 2.

BM and splenic erythropoiesis in 6-week-old WT and Klotho^−/− mice. A: Flow cytometry analysis shows an increase in primitive Pro-Es (Ter119^med CD71^high) in Klotho^−/− mice (Pro-E: WT, n = 12; Klotho^−/−, n = 9); inset shows flow cytometry gating. B: Erythroid lineage commitment was analyzed using Ter119 and CD71 markers coupled with FSC measurements. A mature Ery-C (Ter119^high CD71^low FSC^low) population was observed in Klotho^−/− compared with WT littermates (Ery-C: WT, n = 9; Klotho^−/−, n = 7); inset shows flow cytometry gating on the Ter119^high population. C: Colony-forming unit assay showing burst-forming unit–erythroid (BFU-E) colonies produced by Klotho^−/− BM cells (WT, n = 13; Klotho^−/−, n = 11). Cells from each mouse were plated in triplicate, and the number of colonies was scored based on morphologic features. D: Graphic representation of flow cytometry analysis of the mature Ery-C population in Klotho^−/− splenocytes (WT, n = 9; Klotho^−/−, n = 8). E: Colony-forming unit assay showing splenic BFU-E colonies in Klotho^−/− mice (WT, n = 4; Klotho^−/−, n = 4). Cells from WT and Klotho^−/− spleens were plated in triplicate. Data represent the means ± SEM. ^∗P < 0.05 and ^∗∗P < 0.01.