FIGURE 4.

PIP₂- and PIP₃-binding domains do not target GFP to sites of vesicle budding or to EMVs. A–H, fluorescence and phase microscopy images of N-Rh-PE-labeled Jurkat T-cells expressing GFP-SYN (A–D) or AKT-GFP (E–H). Scale bar, 10 μm. I, anti-GFP immunoblot of EMV and cell lysates prepared from Jurkat T-cells expressing AKT-GFP, AKT-TyA-GFP, GFP-SYN, or TyA-GFP-SYN. The bar graph to the right shows the average ± 1 S.D. of the relative budding calculated from band intensities (EMV/(EMV + cell)) relative to the AcylTyA-GFP control, which were determined by densitometric analysis of immunoblot films. Three asterisks reflect a p value of < 0.0005.