Detection of megakaryocytic markers on Mks derived from CD34⁺ cells in culture. (A) Experimental protocol for the culture conditions used. CD34⁺ cells were isolated from human mobilized peripheral blood samples cultured in the presence of TPO (50 ng/mL) for 4 days, washed, counted, and cultured for up to 12 days. D indicates days from the start of culture. (B) Cultured cells were stained with anti-human CD41-FITC and CD42a-Alexa Fluor 647 simultaneously and analyzed by flow cytometry. Shown is the dot-plot profile of GPIIb and GPIX double-positive Mks on day 8 of culture. (C) Kinetic expression of GPIIb (light gray) and GPIX (dark gray) on Mks on days 4, 7, 8, 9, 11, and 12 of culture. The graph shows the means ± SD.