Figure 1.

Generation and immunoblot analysis of Byg/Tigm compound hets. (A–C) Mating schemes used to generate compound heterozygous mice harboring both the Byg and Tigm mutant Gga2 alleles. The Byg strain was backcrossed for at least eight generations into the C57BL/6 genetic background before being crossed with the Tigm strain, also in the C57BL/6 background. (D and E) Genotyping results and Western blot analysis of brain tissue from day 1 pups of representative litters resulting from the mating schemes shown in (B and C), respectively. Twenty-five μg of protein extract for each sample was subjected to SDS-PAGE and immunoblot analysis of GGA1, GGA2, GGA3, AP-1, and GAPDH (5 μg of lysate) as a control. *A non-specific band. Use of a 4–12% Bris-Tris gradient gel (E) resulted in better separation of the GGA2 band from the non-specific band compared to a 10% Tris-Glycine gel (D).