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Fig. 4. Constitutively active G ${alpha}$ _o rescues inhibition of Wnt3a action by RGS19. (A) F9 cells transiently transfected with vectors containing Fz1, M50 and/or RGS19, Q205L G ${alpha}$ _o, Q209L G ${alpha}$ ₁₁ and Q209L G ${alpha}$ _q were treated with or without Wnt3a for 8 hours, then collected and subjected to the luciferase assay. The results show that expression of Q205L G ${alpha}$ _o (left panel), but not Q209L G ${alpha}$ ₁₁ (right panel) or Q209L G ${alpha}$ _q (lower panel), can rescue the inhibitory effect of RGS19 on Wnt3a-mediated transcription. Results are displayed as the difference in reporter activation upon Wnt3a stimulation (means ± s.e.) as compared with an untransfected control and are representative of three independent experiments. *P<0.05 for the difference between wild-type cells and those expressing either RGS19 or RGS19 and Q209L G ${alpha}$ ₁₁ or Q209L G ${alpha}$ _q. RLU, relative luciferase units. (B) Western blot analysis was used to measure the expression of transiently transfected vectors encoding Q205L G ${alpha}$ _o, Q209L G ${alpha}$ ₁₁ and Q209L G ${alpha}$ _q. Lysates were collected from cells transfected with vectors encoding Q205L G ${alpha}$ _o, Q209L G ${alpha}$ ₁₁ and Q209L G ${alpha}$ _q, then subjected to SDS-PAGE on an 11% acrylamide gel, transferred to nitrocellulose and probed with an antibody against each G-protein.

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