Vasostatin impairs differentiation. (A) Coomassie staining of steps during purification of recombinant Vasostatin; lane 1: whole protein lysates fraction; 2: unbound protein fraction; 3: washing step fraction; and 4: purified and concentrated protein (5 μg protein/lane). Bottom panel indicates Western blot with N-terminus CRT antibody. (B) 0.2 × 10⁶/mL HL-60 cells were treated with 1μM ATRA and/or 2μM vasostatin. CD11b expression was determined by flow cytometry. (C) The proportion of metabolizing cells was assessed via use of the Alamar Blue Assay, and we identified a decrease in metabolizing cells in ATRA-treated HL-60 cells. (D) Cell-cycle analysis (propidium iodide staining) of HL-60 cells treated with ATRA and/or vasostatin.