Fig. 8. MßCD extraction prevents recruitment of ß-arrestin-2-GFP to the plasma membrane by LPA₁ but not by ß₂AR. (A,B) HeLa cells were transiently transfected with plasmid encoding either LPA₁ (A) or ß₂AR (B) along with ß-arrestin-2-GFP. The cells were then left untreated, pre-treated with 5 mM MßCD for 1 hour, or treated sequentially with 5 mM MßCD for 1 hour and 10 mM water-soluble cholesterol for 1 hour prior to incubation with 10 µM LPA for 2 minutes. The cells were then fixed and processed for indirect immunofluorescence microscopy. The inset shows a magnified image of the region of the cell indicated by the arrow. (C) The percentage of cells exhibiting recruitment of ß-arrestin-2-GFP to punctate plasma membrane spots after 2 minutes agonist stimulation was determined by scoring 100 cells per condition for cells expressing LPA₁ and ß₂AR. Bar, 10 µm.