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Schema of the experimental design for identifying early TPA-responsive genes. HL-60 cells were synchronized with nocodazole (0.05 μg/ml for 24 h) and then cells were washed with culture medium and followed by treatment with cycloheximide (CHX, 10 μg/ml) for 30 min prior to subsequent stimulation with TPA (32 nM, 1 h) or DMSO (0.05%, 1 h) as the vehicle control. The cells were then collected and used for microarray and realtime PCR analysis.
