Skip to main content
Figure 3 | BMC Developmental Biology

Figure 3

From: GrlJ, a Dictyostelium GABAB-like receptor with roles in post-aggregation development

Figure 3

Cellular localisation of GrlJ. (A) A GFP-tagged GrlJ fusion protein localises to the plasma membane and intracellular membranes. GrlJ sequences were fused to GFP sequences in such a way that GFP was present at the C-terminus of GrlJ. Shown are grlJ- cells expressing GrlJ-GFP. The cells were fixed with cold methanol and stained for annexinC1 using mAb 185-338-2 [13] or interaptin using mAb 260-60-10 [15]. Detection was with Cy3-labeled secondary antibody. Confocal images are shown. Bar, 10 μm. (B) Distribution of GrlJ-GFP in cell fractionation studies. grlJ- cells were lysed by sonification and the post nuclear supernatant separated into supernatant and pellet by centrifugation at 100.000 g. The proteins were separated by SDS-PAGE (10% acrylamide), the resulting western blot was probed with GFP-specific antibody K3-184-2. α-Actinin, a cytosolic protein, and Comitin, a membrane associated protein, were used for control [14]. L, whole cell lysate, S, 100.000 g supernatant, P, 100.000 g pellet.

Back to article page