American Society of Hematology

Fig. 4.

$Fig. 4. TPO-induced MAPK phosphorylation in purified murine megakaryocytes. Bone marrow cells were harvested from mice preinjected with 2 μg/d human TPO for 5 days, grown in serum-free medium with 35 ng/mL murine TPO for 3 days, and then purified by an albumin density-gradient column. Purified megakaryocytes were incubated in a serum-free, cytokine-free medium for 7 hours and stimulated with 14 ng/mL murine TPO for 10 minutes. Twenty minutes before stimulation, PD 98059 (PD) at 20 μmol/L or 50 μmol/L was added. The cell lysates were size-fractionated, transferred to nitrocellulose, and probed with anti–double-phosphorylated ERK antibody (upper panel). The blot was then stripped and reprobed with anti-ERK2 antibody (lower panel) to assure equal loading in all lanes.$

TPO-induced MAPK phosphorylation in purified murine megakaryocytes. Bone marrow cells were harvested from mice preinjected with 2 μg/d human TPO for 5 days, grown in serum-free medium with 35 ng/mL murine TPO for 3 days, and then purified by an albumin density-gradient column. Purified megakaryocytes were incubated in a serum-free, cytokine-free medium for 7 hours and stimulated with 14 ng/mL murine TPO for 10 minutes. Twenty minutes before stimulation, PD 98059 (PD) at 20 μmol/L or 50 μmol/L was added. The cell lysates were size-fractionated, transferred to nitrocellulose, and probed with anti–double-phosphorylated ERK antibody (upper panel). The blot was then stripped and reprobed with anti-ERK2 antibody (lower panel) to assure equal loading in all lanes.

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