High levels of LSD1 expression and glycolytic activity in erythroleukemia cells. Scatter plots showing positive correlation between LSD1 and GLUT1 expression in AML in TCGA clinical samples (n = 173) (A) and CCLE cell lines (n = 37) (B). Pearson product correlation coefficient and P values are indicated. (C) Glycolysis/OXPHOS balance of HEL and HL60 cells, determined by using an extracellular flux analyzer. Values indicate the ratio of extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Values are mean ± standard deviation (SD) of 10 wells. (D) Expression changes of glycolytic genes in HEL cells expressing short hairpin RNA against LSD1 (shLSD1#1). Full descriptions of gene symbols are provided in supplemental Table 2. qRT-PCR values, which were normalized to the expression levels of the 36B4 gene, are shown as the fold difference against control (ctrl) samples. (E) Decrease of GLUT1 protein was confirmed in LSD1-KD HEL cells. Scanned images of unprocessed blots are shown in supplemental Figure 10. (F) Reduction of glucose uptake in LSD1-KD HEL cells. 2-NBDG incorporation was determined by flow cytometry. Mean fluorescence intensities are shown. (G) Reduced glycolytic activity in LSD1-KD HEL cells. Values are mean ± SD of 5 assays. (H) Expression changes of glycolytic genes under the treatment with the LSD1 inhibitor S2101. (I) Decrease of GLUT1 protein in S2101-treated HEL cells. (J) Reduction of glucose uptake by S2101 treatment. (K) Reduction of glycolytic activity in S2101-treated HEL cells. Values are mean ± SD of 5 assays. All samples were collected at day 4 unless indicated otherwise. All histogram data are mean ± SD of triplicate results unless indicated otherwise. *P < .05, **P < .01 vs control. CMP, common myeloid progenitor; FPKM, fragments per kilobase of transcript per million mapped reads; GMP, granulocyte erythroid progenitor; HSC, hematopoietic stem cell; MEP, megakaryocyte erythroid progenitor.