Fig. 2.

siRNA-mediated arrestin knockdown in ASMCs. ASMCs were transfected with negative control (NC), anti-arrestin2 (AR2), or anti-arrestin3 (AR3) siRNAs using the nucleofection technique (see materials and methods). After 48 h, cells were lysed and arrestin expression levels were determined by Western blotting. A: representative immunoblot showing arrestin2 depletion. B: the same blot is shown after longer exposure to demonstrate arrestin3 (bottom band) depletion. Lane key: NT, nontransfected; AR2, anti-arrestin2 (100 nM); AR3, anti-arrestin3 (100 nM); AR2 + AR3, anti-arrestin2 (100 nM) and anti-arrestin3 (100 nM); NC100, negative control (100 nM); NC200, negative control (200 nM). C: cumulative data showing endogenous siRNA-targeted arrestin depletion. Data are shown as means ± SE for 4 separate transfections undertaken in ASMC preparations from 4 different animals. **P < 0.01 (one-way ANOVA; Dunnett's post hoc test) compared with arrestin expression in nontransfected or NC siRNA-transfected cells.