Figure 4
From: GxcDD, a putative RacGEF, is involved in Dictyostelium development
Cortical localization of the C-terminus of GxcDD containing the ArfGAP-PH tandem and enrichment on phagosomes. (A) Wild type Dictyostelium cells expressing GFP-ArfGAP-PH were fixed and stained with actin specific mAb act1-7, showing complete overlap of the two. (B) Live cell imaging of cells expressing GFP-ArfGAP-PH incubated with TRITC labelled yeast. GFP-ArfGAP-PH enriches at the phagocytic cup (arrow head) and also at the leading edges of the cell (arrow). Pictures captured at different times are shown. (C) Cells expressing GFP-ArfGAP-PH were subjected to Triton X-100 extraction. Supernatant (S) and pellet (P) fractions were separated by 10% SDS-PAGE and the protein immunodetected using GFP specific mAb K3-184-2. GFP-ArfGAP-PH was equally distributed in both the Triton X-100 insoluble cytoskeleton and the soluble fraction.