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Phospholipid binding of PRIP-C2. Lipid overlay assay was performed using a nitrocellulose membrane on which 1 nmol of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylinositol 4,5-bisphosphate (PIP2) were blotted. Each membrane was probed with GST-fused protein indicated on the left followed by immunodetection with anti-GST antibody. GST-fused PRIP-C2, PLCδ-PH, or Syt-C2A were used. Ca2+ (−) indicates incubation with 1 mm EGTA alone.
