Extended Data Figure 7: BCAT1 is required for human myeloid leukaemia. | Nature

Extended Data Figure 7: BCAT1 is required for human myeloid leukaemia.

From: Cancer progression by reprogrammed BCAA metabolism in myeloid leukaemia

Extended Data Figure 7

a, RT–qPCR analysis of BCAT1 expression in the human K562 BC-CML cell line transduced with lentiviral shCtrl or BCAT1 shRNA (shBCAT1-c and shBCAT1-d). The expression levels are normalized and displayed relative to the expression of the B2M control. **P < 0.01. b, Western blot analysis of BCAT1 protein levels in K562 cells infected with the indicated lentiviral shRNA constructs. Human β-tubulin protein (β-Tub) was used as the loading control. β-Tubulin image is the same as shown in Fig. 3j. c, d, Colony-forming ability of (c) K562 cells transduced with control (shCtrl) or BCAT1 shRNAs (shBCAT1-c and shBCAT1-d) and (d) K562 cells cultured with the indicated concentrations of Gbp. One hundred cells were plated per well in triplicate. Photomicrographs show representative colonies formed. Scale bar, 200 μm. Error bars, s.e.m. **P < 0.01, ***P < 0.001. e, RT–qPCR analysis of BCAT1 expression in the samples from the patient with BC-CML used in the data presented in Fig. 3d that were transduced with control (shCtrl) or BCAT1 shRNA (shBCAT1-d). ***P < 0.001. f, Colony-forming ability of primary human CD34+ BC-CML cells from another specimen from a patient treated with Gbp. Error bars, s.e.m. **P < 0.01. gi, Colony-forming ability of (g) MV4-11, (h) U937 and (i) HL60 human AML cells treated with the indicated concentrations of Gbp. MV4-11, HL60 cells (300 per well) or U937 (100 per well) were plated in triplicate. Photomicrographs show representative colonies formed. Scale bars, 200 μm. Error bars, s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001. j, BCAT1 expression in human patients with de novo AML. Data for BCAT1 expression levels from the TCGA AML dataset were divided into quartiles and compared. On average, the top quartile cohort showed 1.6-fold higher expression level than the bottom quartile. **P < 0.01.

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