Fig. 5: Cdk6^KM/KM cells have an increased need for aerobic glycolysis.

A ATP levels determined by CellTiter-Glo® Luminescent Cell Viability Assay; n = 3 cell lines per genotype. Data are depicted as mean ± SD; *p ≤ 0.05. B Cytoplasm/mitochondria ratio of ATP levels as determined by CellTiter-Glo® Luminescent Cell Viability Assay after separation of mitochondria and cytoplasmatic fractions; n = 3 cell lines per genotype in duplicates. Data are depicted as mean ± SD; **p ≤ 0.01. C Pyruvate (left) and lactate (right) levels as determined by high-throughput metabolite analysis in BCR::ABL1+ cell lines; n = 3 cell lines per genotype. Data are depicted as mean ± SD; *p > 0.05. D Extracellular lactate levels measured with Lactate-Glo™ Assay; n = 3 cell lines per genotype; Data are depicted as mean ± SD; *p ≤ 0.05. E Scheme of the inhibitory mechanism of 2-DG. 2-DG and glucose get competitively transported into the cell where they get both phosphorylated in the first step of glycolysis. However, 2-DG-6-P cannot be further metabolized, accumulates within the cell and inhibits both up- and downstream enzymes of glycolysis. F AnnexinV/7AAD stainings of BCR::ABL1+ cells on day 13 of a proliferation experiment (see Supplementary Fig. S5E); n = 3 cell lines per genotype. Data are depicted as mean ± SD; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. G AnnexinV/7AAD stainings of human BCR::ABL1+ SUPB15 cells on day 14 of proliferation experiment (see Supplementary Fig. S5H); n = 3 replicates. Data are depicted as mean ± SD; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. H Number of living HPC^LSK cells compared to murine BCR::ABL1+ cells in a co-culture setting with the indicated treatment at day 10; n = 3 replicates. Data are depicted as mean ± SD. HPC^LSK fraction relative to BCR::ABL1+ was compared between treatment conditions; **p ≤ 0.01, ns not significant.