After 24 h exposure to bortezomib (10 nM), carfilzomib (5 nM), ixazomib (50 nM), alisertib, volasertib (500 nM) and avapritinib (250 nM) alone or in combination with bortezomib or alisertib, apoptotic cell death was assessed in HMC-1 and ROSAKIT D816V cells by using the Annexin-V-FLUOS Staining Kit by Roche Applied Science, according manufacturer instructions and measuring the uptake of fluorescinated Annexin V and propidium iodide (both from Roche). A FACSCanto II flow cytometer (Beckton Dickinson) set at 488 nm excitation and 530 nm wavelength bandpass filter for fluorescein detection or 580 nm for PI detection and a dedicated software (DIVA, Beckton Dickinson) were used for analysis.
Mancini M., Monaldi C., De Santis S., Papayannidis C., Rondoni M., Sartor C., Bruno S., Pagano L., Criscuolo M., Zanotti R., Bonifacio M., Tosi P., Arock M., Valent P., Cavo M, & Soverini S. (2023). SETD2 non genomic loss of function in advanced systemic mastocytosis is mediated by an Aurora kinase A/MDM2 axis and can be therapeutically targeted. Biomarker Research, 11, 29.
Corresponding Organization : Istituto di Ematologia di Bologna
Other organizations :
University of Bologna, Azienda Unità Sanitaria Locale Della Romagna, Università Cattolica del Sacro Cuore, Agostino Gemelli University Polyclinic, Istituti di Ricovero e Cura a Carattere Scientifico, University of Verona, Ospedale Infermi di Rimini, Pitié-Salpêtrière Hospital, Sorbonne Université, Medical University of Vienna
Annexin-V-FLUOS Staining Kit by Roche Applied Science
FACSCanto II flow cytometer (Beckton Dickinson) set at 488 nm excitation and 530 nm wavelength bandpass filter for fluorescein detection or 580 nm for PI detection
DIVA software (Beckton Dickinson) for analysis
positive controls
Bortezomib or alisertib (in combination with other treatments)
Annotations
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