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. 2009 Aug;23(8):1272–1280. doi: 10.1210/me.2009-0047

Fig. 4.

Fig. 4.

siRNA-mediated depletion of endogenous GRKs in ULTR and primary myometrial cells. Cells were transfected with siRNA constructs targeting endogenous GRK2, GRK3, GRK5, and GRK6 as described in Materials and Methods. After 48 h cells were lysed and 40 μg of protein loaded per lane for SDS-PAGE separation and immunoblotting. Representative immunoblots showing effects of anti-GRK siRNA treatment on endogenous GRK expression in ULTR (A) and primary myometrial (B) cells: lane 1, negative control; lane 2, anti-GRK2; lane 3, anti-GRK3; lane 4, anti-GRK5; lane 5, anti-GRK6. Densitometric analysis was undertaken on all blots and data are shown to highlight changes in GRK2, GRK3, GRK5, and GRK6 expression in (C) ULTR or (D) primary myometrial cells after anti-GRK2, anti-GRK3, anti-GRK5, or anti-GRK6 siRNA treatments. Data are shown as means ± sem for three to six separate experiments in which GRK isoenzyme expression was compared with cells transfected with a negative control siRNA (=100% for each GRK isoenzyme); **, P < 0.01 comparing anti-GRK2/anti-GRK3/anti-GRK5/anti-GRK6 siRNA treatments with respective negative control siRNA treatments.