Fig. 3.

Ligand- and shear stress-induced dephosphorylation of β-arrestin-1. HCAECs were treated with S1P (2.5 μM) or exposed to flow (14 dyn/cm²) for indicated time points. Immunoblotting was performed on cell lysates using antibodies against phosphorylated β-arrestin-1 (S412) and total β-arrestin-1 to assess the level of phosphorylation/dephosphorylation. As a positive control for cell stimulation, lysates were also probed for phosphorylated Akt (S473) and total Akt. Representative blots from 5 independent experiments are shown. Bar graph represents the quantification as a ratio of phospho-S412 over total β-arrestin-1, with the mean value of the control condition arbitrarily set to 100% and error bars indicating SE; n = 5. *P < 0.05.