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Assay for intramolecular interactions. Cell lysates expressing PTEN-GFP or PTENA4-GFP are incubated with cell lysates expressing PTEN352–403-YFP-FLAG. Introducing the A4 mutation into full length PTEN allowed interactions of the core region of PTENA4 and the FLAG-tagged C-terminal tail. PTEN352–403-YFP-FLAG was immunoprecipitated with beads coupled to anti-FLAG antibodies. Bound fractions (IP) are analyzed with antibodies to GFP and FLAG.
