Figure 2. Evaluation of GRK2 expression and subcellular localization in response to BK.
After stimulation with BK (30 nM) for 5 and 15 min, BAEC were collected and processed to isolate cellular compartments or to obtain whole cell lysate where GRK2 level was evaluated by western blot. In whole cell lysate, GRK2 increased at 5 min from BK stimulation and returned to baseline levels at 15 min (A). In membrane extract, GRK2 accumulates at 5 min from BK stimulation and continued to be higher at 15 min (B). Mitochondrial and cytosolic levels of GRK2 increased at 5 min from BK stimulation, reflecting the levels of GRK2 in whole cell lysate (C-D). Actin was used as a loading control for whole cell lysate and for cytosolic extracts, while Gαi and VDAC were used as loading control for membrane and mitochondrial extracts, respectively. Images are representative of at least three independent experiments (One-way ANOVA: *p<0.05, n≥3).