grin2bb transcript and protein levels are unaffected in bigheart. (A) pGBT-PX integration in bigheart maps to intron 2 of grin2bb gene located on the reverse strand of Chromosome 1. Box in red indicates the 5′RACE mapped region, blue represents the 3′ RACE mapped region, and the green represents the iPCR mapped region. (B) Screen shot of UCSC genome bowser showing the grin2bb transcript and the bigheart GBT insertion loci in the intron 2. (C) Expression profiling of grin2bb transcript in 9-month-old adult WT heart tissue using in situ hybridization. (D) qRT PCR showing relative expression of grin2bb partial transcript in sibling wildtype and bh−/− heart tissue. (E) Western blot analysis of grin2bb in WT and bh−/− heart tissue. β-actin was used as internal control. (F) Quantification showing Grin2bb protein fold change in the WT and bh−/−. (black arrows in A indicate the regions where the primers were designed. The RT PCR and western blot experiments were repeated three times. The bar indicates the average values ± SD.
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