Effects of Lsd1 deletion in erythroid cells of CKO mice.Lsd1 CKO and control mice were administered 7 intraperitoneal injections of Tx on alternate days. Total BM cells were harvested and processed for colony assays and flow cytometric analyses. (A) Colony numbers of CFU-GEMM (GEMM), CFU-GM (GM), BFU-E, and CFU-E cells per 10⁵ BM cells from control and Lsd1 CKO mice. (B) Representative micrographs of BFU-E from control or Lsd1 CKO mice taken at low or high (inset) magnification on day 10 of the CFU assay. Scale bars, 200 μM. (C) No statistically significant difference in the spleens (as percentages of body weights [wt.]) was observed between control and Lsd1 CKO mice. (D) Representative flow cytometry plots displaying the gating for CFU-E cells from control and Lsd1 CKO mice. (E) Bar graph showing absolute numbers of CFU-E cells in control and Lsd1 CKO mice (per 2 femurs + 2 tibias). (F) Representative flow cytometric plots showing the gating strategy for defining CD71⁺Ter119⁺ erythroid precursor cells (upper panels) that were subsequently gated by CD44 staining vs forward scatter (FSC) to separate BasoEs, PolyEs, and OrthoEs (lower panels). (G) Absolute numbers of BasoEs, PolyEs, and OrthoEs in control and Lsd1 CKO BM. (H) Deletion efficiency of Lsd1 floxed alleles in flow-sorted BasoEs was determined by qPCR. Primer pair P1 detects Lsd1 genomic DNA flanked by the 2 loxP sites, whereas primer pair P2 detects Lsd1 genomic DNA that is unaffected by Cre-mediated deletion. Data are mean ± standard deviation. *P < .05; **P < .01; ***P < .001, unpaired Student t test.