Fig. 3: Molecular mechanisms of cabozantinib-induced AML1-ETO downregulation.
From: Repurposing cabozantinib with therapeutic potential in KIT-driven t(8;21) acute myeloid leukaemias
A Kasumi-1 cells were treated with DMSO or various concentrations of cabozantinib for 24 h, 48 h, and 72 h before harvesting. Western blot analysis of the time course of changes in protein levels of AML1-ETO. B Western blot analysis of the time course of changes in the protein level of AML1-ETO after DMSO (control), CHX alone or in combination with cabozantinib. Signal intensity was quantified with ImageJ and normalised to β-actin and control (0-h) group. C The protein expression of AML1-ETO was measured in Kasumi-1 cells, which were treated with 500 nM cabozantinib for indicated hours before harvested for western blot analysis. The proteasome inhibitor MG-132 (5 μM) was added 4 h before cell harvest. D Monitoring protein synthesis in Kasumi-1 and OCI-AML3 cells by puromycin labelling. β-actin was loaded as a control. Signal intensity was quantified with ImageJ and normalised to β-actin and DMSO controls. E Co-immunoprecipitation (co-IP) followed puromycin-labelled de novo synthesis protein showed decreased AML1-ETO protein expression level. Mouse IgG was used as a negative control of co-IP.
