May grunwald and giemsa

A minimum of two biological replicates were evaluated for each assay. Dose response curves were generated using a 4-parameter equation (XLfit, IDBS). All kits were used according to the manufacturer’s recommendations. Cell cycle phase distribution was determined by flow cytometry using the Cycletest™ PLUS DNA Reagent Kit (BD Biosciences) and data were analyzed using FlowJo software (TreeStar Inc.). Proliferation was assessed using BrdU Cell Proliferation Assay Kit (Cell Signaling Technology) with a 4-hour pulse. Values were expressed as percent of vehicle. Caspase activity was measured using Caspase-Glo 3/7 (Promega). Luminescence values were normalized to CTG (Promega) for cell number. Peak activation was determined for MOLM-13, OCI-AML3, MV-4-11, THP-1, SIG-M5, HL-60, and Kasumi-1 to be days 4, 6, 6, 3, 4, 5, and 5, respectively. Values were expressed as a fold change relative to vehicle. Gene expression was evaluated using real time-quantitative polymerase chain reaction (RT-qPCR), as previously described.^{11 (link)} Superoxide anion production was measured using LumiMax Superoxide Anion Detection Kit (Agilent Technologies). Values were expressed as fold increase over vehicle. Morphology was visually assessed after staining with May-Grunwald and Giemsa (Sigma).