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Fig. 2. The PLD1 PX domain interacts with PtdIns(3,4,5)P3 in NIH-3T3 cells. NIH-3T3 cells grown on cover slips were transfected with pEGFP-PLD1 PX, pEGFP-PLD1 PX(R179K), pEGFP-PLD1 PX(R179A), or pEGFP-Btk PH. (A) After 24 hours of serum starvation, cells were treated with PDGF (25 ng/ml). After 5 minutes, cells were fixed and the localization of PLD1 PX domains was visualized by confocal microscopy. Bars, 10 µm. (B) Cells as described in (A) were cultured in dishes and fractionated into cytosolic (Cyt) and membrane (Mem) fractions. Western blotting was performed with anti-EGFP antibody and show that only the wild-type PLD1 PX domain showed membrane localization after PDGF treatment.





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