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. 2005 Oct 10;102(42):15030–15035. doi: 10.1073/pnas.0507533102

Fig. 3.

Fig. 3.

Role of O2, UBR1, and UBR2 in proteolysis of RGS4 and RGS5. (A) RGS5 and C2A-RGS5 were coexpressed with luciferase in reticulocyte lysates under a normoxic or hypoxic condition, followed by anti-RGS5 or biotin-based Western blotting. (B Upper) RGS5 and C2A-RGS5 were expressed in reticulocyte lysates in the presence of MG132 either under a normoxic or hypoxic condition, followed by anti-Ub immunoprecipitation and a subsequent antibiotin Western blotting. (B) (Bottom) Comparison of RGS5 expression using anti-biotin Western blotting. (C) RGS5 was coexpressed with LacZ in +/+ and ATE1–/– EFs in normoxic and hypoxic (0.1% O2) condition, followed by anti-RGS5, LacZ, and actin immunoblotting. (D) Pulse–chase analysis of RGS4 and RGS5 in +/+, UBR1–/–, UBR2–/–, and UBR1–/–UBR2–/– cells. The transfected cells were labeled for 12 min with [35S]Met/[35S]Cys, followed by anti-RGS4 or RGS5 immunoprecipitation, SDS/PAGE analysis, and autoradiography. (E) Quantitation of data in D using PhosphorImager.