Figure 1.

Targeted disruption of the murine Gα_q gene. (A) Part of the wild-type Gα_q locus containing the two last exons (wt allele), the targeting construct, and the targeted locus (mut. allele) are shown. Neo, neomycin resistance gene. The sizes of the AflII fragments predicted to hybridize to the indicated diagnostic probe are shown. Restriction endonucleases: A, AflII; B, BglII; N, NdeI; S, SacI; Sm, SmaI; and X, XhoI. (B) Southern blot analysis of AflII-digested genomic DNA from wild-type (+/+), hemizygous (−/+), and homozygous mutant mice (−/−) with the diagnostic probe indicated in A. (C) Western blot analysis of whole brain cholate extracts from wild-type (+/+) and homozygous Gα_q mutant mice (−/−) with antibodies recognizing the α-subunits of G_q (Gα_q), G₁₁ (Gα₁₁), G₁₃ (Gα₁₃), and G_o (Gα_o). (D) Western blot analysis of cerebellar membrane fractions from wild-type (+/+) and homozygous Gα_q mutant animals (−/−) with antibodies recognizing the α-subunits of G_q and G₁₁ (Gα_q/Gα₁₁), G_o (Gα_o), type 1 metabotropic glutamate receptor (mGluR1), phospholipase C-β3 (PLC-β3), and phospholipase C-β4 (PLC-β4).