Figure 3.

The hydrophobic amino acids involved in the interdomain association of PLD1 are required for its vesicular localization. (A) Alignment of the HKD motifs in mammalian PLD1 from different species and schematic representation of mutations generated in the GFP-PLD1. (B) HEK 293 cells were transfected with the indicated GFP-PLD1 constructs and its localization was observed using fluorescence microscopy. WT, wild type (C) HEK 293 cells were transfected with the indicated GFP-PLD1 constructs and the lysates were fractionated into cytosol (C) and vesicles (V), after which the samples weresubjected to immunoblotanalysis with the indicated antibodies. These data are representative of results obtained from three experiments.