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Lysates from 293T cells transfected with wild-type SRPK1 and its mutant forms were analyzed under non-reducing conditions (i.e., without DTT) or reducing conditions (in the presence of 90 mM DTT) on 10% SDS-polyacrylamide gels. The proteins were then transferred to nitrocellulose and epitope-tagged wild-type or mutant SRPK1 was detected with the M5 anti-FLAG monoclonal antibody.
