Figure 5.
Leukemic cells are dependent on an exogenous source of Ser/Gly (A) Effect of Ser and/or Gly starvation on leukemic cells. MLL-ENL–transduced primary HSPCs were incubated in minimal essential medium supplemented with the indicated concentrations of Ser and Gly or a combination of both amino acids. Proliferation was recorded by standard MTT assays. (B) Selective starvation for Ser and Gly reduces the number of leukemic CFCs. Mean and standard deviation of the relative number of CFCs obtained after a transient starvation of normal and leukemic HSPCs for Ser and Gly. This experiment was performed in triplicate. (C) A sample aspect of CFC colonies in methylcellulose obtained with normal and leukemic cells grown for 48 hours, with or without Ser/Gly. Note that the methylcellulose medium used to enumerate CFCs contains standard amounts of the amino acids. (D) ROS production in normal and leukemic cells after amino acid starvation, as above. FACS plots after staining with CellROX deep-red reagent are shown. (E) Cell cycle distribution of cells as in panel D. (F) Surface staining of leukemic cells for the differentiation marker Gr-1 (Ly6-G) cultivated for 48 hours in supplemented or Ser/Gly-starved conditions. (G) Effect of Ser/Gly deprivation on CFC capacity of MLL-r samples from patients. Cells obtained from leukemic blasts of 2 patients diagnosed with an MLL-AF4 and an MLL-AF6 translocation respectively were cultivated for 48 hours in medium, with or without Ser/Gly, and subsequently plated in duplicate into fully supplemented methylcellulose medium. Colonies were stained and photographed. (H) Survival curve of leukemia cell transplant-recipient animals under different nutrient conditions. Transplant recipients grafted with syngenic MLL-ENL–transduced cells were separated into 2 cohorts fed with either complete chow as the control (green line) or with specifically formulated Ser/Gly-free diet that was otherwise of identical composition (red line). Given are the mean and standard deviation of mean survival times. (I) Spleen weight of animals as recorded in panel H.

Leukemic cells are dependent on an exogenous source of Ser/Gly (A) Effect of Ser and/or Gly starvation on leukemic cells. MLL-ENL–transduced primary HSPCs were incubated in minimal essential medium supplemented with the indicated concentrations of Ser and Gly or a combination of both amino acids. Proliferation was recorded by standard MTT assays. (B) Selective starvation for Ser and Gly reduces the number of leukemic CFCs. Mean and standard deviation of the relative number of CFCs obtained after a transient starvation of normal and leukemic HSPCs for Ser and Gly. This experiment was performed in triplicate. (C) A sample aspect of CFC colonies in methylcellulose obtained with normal and leukemic cells grown for 48 hours, with or without Ser/Gly. Note that the methylcellulose medium used to enumerate CFCs contains standard amounts of the amino acids. (D) ROS production in normal and leukemic cells after amino acid starvation, as above. FACS plots after staining with CellROX deep-red reagent are shown. (E) Cell cycle distribution of cells as in panel D. (F) Surface staining of leukemic cells for the differentiation marker Gr-1 (Ly6-G) cultivated for 48 hours in supplemented or Ser/Gly-starved conditions. (G) Effect of Ser/Gly deprivation on CFC capacity of MLL-r samples from patients. Cells obtained from leukemic blasts of 2 patients diagnosed with an MLL-AF4 and an MLL-AF6 translocation respectively were cultivated for 48 hours in medium, with or without Ser/Gly, and subsequently plated in duplicate into fully supplemented methylcellulose medium. Colonies were stained and photographed. (H) Survival curve of leukemia cell transplant-recipient animals under different nutrient conditions. Transplant recipients grafted with syngenic MLL-ENL–transduced cells were separated into 2 cohorts fed with either complete chow as the control (green line) or with specifically formulated Ser/Gly-free diet that was otherwise of identical composition (red line). Given are the mean and standard deviation of mean survival times. (I) Spleen weight of animals as recorded in panel H.

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