Figure 8. L148S interacts with β-arrestin-1 and 2 while GPR54 continues to modulate gene expression in the absence of G_q/11.

(A) Representative autoradiograph showing the co-immunoprecipitation of β-arrestin-1 and β-arrestin-2 with FLAG-L148S. HEK 293 cells were transfected with 10 µg of β-arrestin-1 or -2-GFP and 10 µg of FLAG-L148S or empty FLAG vector. Cells were left untreated or stimulated with 100 nM Kp-10 for 5 minutes. Lysates were prepared, immunoprecipitated with mouse anti-FLAG antibody, and immunoblotted with mouse anti-GFP antibody. The expression of β-arrestin-1 or -2-GFP in 50 µg of total protein from the corresponding HEK 293 cell lysates is also shown. The data represent the mean ± SE for three independent experiments. IB: Immunoblotting; IP: immunoprecipitation. (B) Gene microarray analysis of G_q/11 KO MEFs versus WT control MEFs that were treated with Kp-10 for six hours. The data is presented as a three-way Venn diagram and shows the overlap of genes changing greater than 2 fold in their expression (up or down) as a proportion of all the probesets on the Affymetrix Mouse Gene 1.0 ST Array. In total, 1,635 out of a total of 35,556 probesets are altered greater than 2-fold (4.6%) (P<0.05).