Figure 5.

Expression of Gα_t-2, Gα_gust, and members of the T2R family in STC-1 cells. (A) RT-PCR analysis for the α subunits of G_t-₂ and G_gust was performed on poly(A)⁺ RNA isolated from STC-1 cells. PCR products with the predicted size were subcloned and sequenced to confirm their identity. (B) Immunoblot analysis for Gα_t-2 and Gα_gust was performed on total protein extracts prepared from STC-1 cells. Normal or preabsorbed Gα_t-₂ and Gα_gust-specific antibodies were used to detect the presence of these Gα subunits in STC-1 cell lysates by Western blotting. (C) RT-PCR analysis using rat T2R subtype-specific primers was performed on the same cDNAs used in the experiments described in A. PCR products corresponding to the predicted mT2Rs were subcloned and sequenced to confirm their relatedness to published rat and mouse sequences.