Double-stranded siRNA duplexes with 3′-dTdT overhangs were obtained from Qiagen for GRK2 (5′-AAGAAAUUCAUUGAGAGCGAU-3′), GRK3 (5′-AAGCAAGCUGUAGAACACGUA-3′), and a nonsilencing RNA duplex (5′-GCUUAGGAGCAUUAGUAAA-3′). HEK293 cells were transfected with 150 nM siRNA for single transfection or with 100 nM of each siRNA for double transfection using HiPerFect (Qiagen). Silencing was quantified after 3 days by performing a GRK β-Gal complementation assay. All experiments showed that the siRNA reduced the target protein levels by ≥80% (fig. S4E).

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