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Fig. 6 | Journal of Hematology & Oncology

Fig. 6

From: RUNX1 mutations promote leukemogenesis of myeloid malignancies in ASXL1-mutated leukemia

Fig. 6

RUNX1-R135T mutation interacted more with HIF-1α and enhanced HIF-1α expression in ASXL1-R693X-mutated leukemia cells. a Transiently expressed FLAG-tagged RUNX1-WT, RUNX1-R135T, and EV control in HEK293T cells or stable-expressed in K562 cells treated with 100 μM CoCl2 for 24 h; immunoprecipitation was performed using anti-FLAG M2 affinity gel, and immunoblot was performed using HIF-1α, RUNX1, and FLAG antibody. β-actin representing as a loading control. b After transient expression of FLAG-tagged WT and mutant RUNX1 in HEK293T cells, cycloheximide (CHX) (100 μg/mL) was treated for different times. Cell lysates were immunoblotted with antibodies against indicated proteins. c, d After stable expression of RUNX1-WT and RUNX1-R135T in K562 cells (c) or ASXL1-R693X, RUNX1-R135T and coexisted expression of ASXL1-R693X and RUNX1-R135T including control in U937 cells (d) were treated with 100 μM CoCl2 or no CoCl2 for 24 h and checked HIF-1α expression by immunoblot analyses. The values in the immunoblot data indicating relative signal density corresponding to Actin or GAPDH expression. All signals from immunoprecipitation were standardized relative to the signal from EV control

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