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The mutant PLD1 proteins are expressed at wild-type levels and exhibit wild-type membrane association. HEK293 cells overexpressing wild-type and mutant PLD1 alleles were lysed and assayed using Western blot analysis. The PLD1 proteins were detected using a rat anti-HA tag monoclonal antibody. Wild-type PLD1 migrates as a doublet; the more slowly migrating band results from phosphorylation, which occurs only when PLD is membrane associated (Kim et al., 1999, 2000; Zhang et al., 1999).
