Figure S1.
Diagrams of methods used in this study.(A) Fluorescence biosensor method. GFP-PHPLCδ1 expressed in living cells binds to PtdIns(4,5)P2 in cellular membranes. (B) Unroofed membrane method. Culture cells on a substrate are stripped of the dorsal plasma membrane and the cytoplasm, and the basal plasma membrane remaining on the substrate is chemically fixed and labeled for PtdIns(4,5)P2. (C) SDS-FRL of quick-frozen cells. Cells are quick-frozen and freeze-fractured to prepare replicas. The membrane physically stabilized by the platinum and carbon coating is labeled for PtdIns(4,5)P2 after SDS treatment. In the present study, some of the replicas before labeling are further treated with protease K to digest integral membrane proteins.

Diagrams of methods used in this study. (A) Fluorescence biosensor method. GFP-PHPLCδ1 expressed in living cells binds to PtdIns(4,5)P2 in cellular membranes. (B) Unroofed membrane method. Culture cells on a substrate are stripped of the dorsal plasma membrane and the cytoplasm, and the basal plasma membrane remaining on the substrate is chemically fixed and labeled for PtdIns(4,5)P2. (C) SDS-FRL of quick-frozen cells. Cells are quick-frozen and freeze-fractured to prepare replicas. The membrane physically stabilized by the platinum and carbon coating is labeled for PtdIns(4,5)P2 after SDS treatment. In the present study, some of the replicas before labeling are further treated with protease K to digest integral membrane proteins.

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