Figure 5
Translocation responses of the Tubby domain and PLCδ1PH domain after rapid dephosphorylation of PtdIns(4,5)P 2 at the PM. COS-7 cells were co-transfected with the PM-targeted FRB tagged with CFP (not shown), the type-IV 5-ptase domain tagged with cerulean (not shown), the GFP-Tubby-domain and PLCδ1PH-mRFP. After 24 h, live cells were analyzed by confocal microscopy. Addition of 100 nM rapamycin induces translocation of the 5-ptase to the PM, causing a complete loss of both PtdIns(4,5)P2 reporter localization. Panel A shows representative images before and after rapamycin addition. Panel B shows a full time course of cytoplasmic fluorescence intensity changes based on the average of 10 cells (Mean ± S.E.M.) expressed as a -fold increase over initial values (upper) or as a percent of the maximal response (lower panel, means are shown only).