Generation of type 1 diabetic Akita mice with cardiac specific conditional knockout of GSK3β.

A: Schematic outline describing the targeting strategy and predicted fragments for generating GSK3β mice with a floxed allele in exon 2 crossed with mice expressing a MerCreMer recombinase under the control of the cardiac-specific αMHC promoter. B. PCR analysis. Genomic DNA was isolated from the atria and analyzed by PCR for tamoxifen induced Cre-mediated excision of GSK3β using primers to the region flanking the floxed allele which gave bands at 885bp, 1,095bp and 250bp respectively. PCR (upper panel) demonstrated that the GSK3β floxed allele is excised only in the Cre+/tamoxifen injected mice. The bottom panel displays genomic detection of the Cre transgene. C: Representative immunoblots of GSK3β from atria in WT, Akita, placebo and tamoxifen treated Akita GSK3βfl/flCre+ mice. GAPDH was used as a loading control. D: Bar graph of data in C normalized to GAPDH. Results are reported as mean ± SEM, *P<0.05, N = 5 for each.