Ba/F3 Cell Proliferation and Viability Assays

Cell number and viability were determined using a hemocytometer by Trypan blue staining. As an assessment of cytotoxicity, Ba/F3 cells were seeded at a density of 1,000 cells per well in 96-well culture plates with 200 μL of media per well. Different concentrations of avadomide or ponatinib were printed in six biological replicates using the Tecan D300e Digital Dispenser. After 48 hours of drug treatment, total cellular ATP content was measured using CellTiter-Glo Luminescent Cell Viability Assay (Promega) according to the manufacturer protocol. Luminescence was assessed by an Envision Microplate Reader (PerkinElmer). Results were normalized to the vehicle control.