Supplementary Methods and Supplementary Figures 1 through 18 from Chemotherapy-Resistant Human Acute Myeloid Leukemia Cells Are Not Enriched for Leukemic Stem Cells but Require Oxidative Metabolism

Supplementary Methods. Supplementary Figure S1. Clinical distributions of AML patients from TUH and 3 independent patient cohorts. Supplementary Figure S2. In vivo treatment with 60 mg/kg/d of cytarabine (AraC) given daily for 5 days induces a significant reduction of the total cell tumor burden in AML-engrafted mice. Supplementary Figure S3. In vivo cytarabine (AraC) treatment induces significant but heterogeneous response in bone marrow and spleen of AML-xenografted NSG mice. Supplementary Figure S4. Comparative analysis of the in vivo response to cytarabine (AraC) with clinicobiological data of AML patients. Supplementary Figure S5. In vivo cytarabine (AraC) treatment induces changes in CD34+/-CD38+/- phenotypes in AML-engrafted mice. Supplementary Figure S6. In vivo cytarabine (AraC) chemotherapy treatment leads to reduction of the absolute number of human CD34+CD38+/- populations in AML. Supplementary figure S7. No enrichment in G0 quiescent cells was observed in mice treated with sublethal dose of cytarabine (AraC) for 5 days in vivo. Supplementary figure S8. Mitochondrial and energetic features of LOW (KG1, KG1a, U937) and HIGH (MOLM14, MV4-11, HL60) OXPHOS AML cell lines. Supplementary figure S9. Functional analysis of the transcriptomes of LOW (KG1a, U937) versus HIGH (MOLM14, HL60) OXPHOS AML cell lines untreated or treated with metformin. Supplementary figure S10. AML cells surviving after cytarabine (AraC) treatment are resistant to chemotherapies and are pre-existing CD36+CD44+ phenotype with an increased oxidative metabolism. Supplementary figure S11. Culture in galactose induces energetic shift of LOW OXPHOS AML U937 cells toward HIGH OXPHOS state, leading to cytarabine (AraC) resistance in AML. Supplementary figure S12. Energetic shift of mtDNA-depleted Rho0 MOLM14 cells toward LOW OXPHOS state induces AraC sensitivity. Supplementary figure S13. Electron Transfer Chain Complex I inhibition by Phenformin (Phenf) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S14. Electron Transfer Chain Complex I inhibition by Metformin (Met) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S15. Electron Transfer Chain Complex I inhibition by Rotenone (Rot) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S16. Electron Transfer Chain Complex III inhibition by Antimycin A (AntiA) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S17. Electron Transfer Chain Complex III inhibition by Atovaquone (ATQ) induces energetic shift toward LOW OXPHOS state and increases cytarabine (AraC) sensitivity in MOLM14 cells. Supplementary figure S18. Working model of the resistance to AraC in vivo.