The cells were cultured at a constant optimal seeding density in Human Methylcellulose Base Media (R&D Systems, Minneapolis, MN, USA) in a 12-well plate to yield colony outgrowth of 20–100 colonies per well as described previously [51 (link),52 (link)]. To test the clonogenic potential, the cells were cultured in the presence of increasing concentrations of DJ4 (0–10 μM) or the DMSO vehicle in a methylcellulose medium for 7–14 days. Blast colonies (>20 cells/colony) were counted under a light microscope and imaged with an Olympus CKX31 inverted microscope (Olympus Corporation, Center Valley, PA, USA) using a 4× objective.
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