Nocodazole and phorbol ester act synergistically on maturation of megakaryocytes. (A) Schematic presentation of the culture conditions. The details are as follows. [1] UT-7 cells were cultured for 7 days in the presence of 0.1% DMSO, which was used as a solvent for nocodazole and TPA. [2] The cells were cultured for 2 days with DMSO. TPA was then added at the concentration of 10 ng/mL, and the culture was subsequently continued for 5 days. [3] The cells were treated with 50 ng/mL of nocodazole for 2 days, and the medium was replaced with those containing 0.1% DMSO. The culture was continued for additional 5 days. [4] The cells were treated with 50 ng/mL of nocodazole for 2 days, and the medium was replaced with those containing 10 ng/mL TPA. The culture was continued for an additional 5 days. (B) Wright-Giemsa staining of UT-7 cells was performed after the culture with four different conditions as shown above. No morphologic change was noted in the condition [1] (not shown). (Original magnification × 400). (C) The intracellular concentration of β-TG was determined by ELISA after the culture in each condition. The mean ± SD (bar) of three independent experiments is shown.