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Activating mutations decreased the ability of a JAK2 fragment encompassing the SH2 and pseudokinase domains to inhibit JAK2 kinase domain. HA-tagged wild-type or mutant fragments were expressed at two amounts (0.5 and 1 μg of DNA) with V5-tagged JAK2 kinase domain in HEK293T cells. JAK2 kinase activity was determined using anti-phospho-JAK2 antibodies. V, vector.
