Erythroid progenitors in the BM and spleen are reduced in 5-FU-treated ESAM-KO mice.

<p>(A-I) WT and ESAM-KO mice were treated with 150 mg/kg of 5-FU, and FACS analyses or methylcellulose cultures were performed at day 8. Then, the number of myeloid and erythroid progenitors from BM, spleen, or PB was evaluated. (A) The number of CMPs, GMPs, and MEPs in BM is shown (n = 4 in each). (B-C) BM cells and splenocytes were subjected to methylcellulose colony formation assays for counting BFU-E and CFU-E. Each bar represents the number of BFU-E (left panel) or CFU-E (right panel) in BM (B) and spleen (C) (n = 6 in each). (D-E) In the left panels, representative FACS profiles of BM (D) and spleen (E). In the right panels, quantification of c-Kit^- Ter119⁺ CD71^Hi cells in BM (D) and spleen (E) (BM; n = 7 in each, spleen; n = 6 in each). (F-H) CMPs, GMPs, and MEPs were sorted from 5-FU-treated BM of WT or ESAM-KO mice. Then, 1 × 10³ MEPs, 2× 10² CMPs, or 1 × 10³ GMPs were subjected to methylcellulose colony formation assays for counting BFU-E (F), CFU-Mix (G), or CFU-G/M/GM (H), respectively (n = 3 in each). (I) The numbers of BUF-E in nucleated cells derived from 100 μL of PB are shown (n = 5 in each). (A-I) The blue bars represent the results for the WT mice, and the red bars represent those of ESAM-KO mice. All data are from one of two independent experiments that gave similar results. Data are shown as mean ± SEM. Statistically significant differences are represented by asterisks (*<i>P</i> < 0.05, ** <i>P</i> < 0.01, *** <i>P</i> < 0.001).