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. Author manuscript; available in PMC: 2010 Feb 1.
Published in final edited form as: Biochim Biophys Acta. 2008 Oct 8;1793(2):406–417. doi: 10.1016/j.bbamcr.2008.09.011

Figure 6. Enhanced receptor phosphorylation and ß-arrestin1/2 membrane translocation correlate with increased endocytosis.

Figure 6

Figure 6

A) Flow cytometric analysis of endocytosed FPR. Cells were incubated for 15 min at 37°C in the presence or absence of 100 nM fMLF, washed to remove remaining ligand, and incubated with a fluorescent formylated hexapeptide to detect FPR remaining on the cell surface. The data are shown as percentage of binding sites on the cell surface, compared to cells incubated in the absence of fMLF. Data show means ± SD from a minimum of three different experiments. P = 0.0057 (non-parametric one-way ANOVA). B) Immunofluorescence analysis of total FPR after 0 or 10 min incubation with fMLF. Methanol fixed and permeabilized cells were incubated with mAb NFPR1 and fluorescent secondary antibody to stain total FPR. (Results for FPR Δ309–315 are not shown since mAb NFPR1 does not bind the mutant receptor.) C) Immunofluorescence analysis of non-phosphorylated FPR after 0, 10 or 60 min incubation with fMLF. Methanol fixed and permeabilized cells were incubated with mAb NFPR2 and fluorescent secondary antibody to stain non-phosphorylated FPR. Bar 50 µm.