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. 2013 Aug 12;8(8):e72173. doi: 10.1371/journal.pone.0072173

Figure 1. Generation of transgenic mice overexpressing Pgam2 in a heart-specific manner.

Figure 1

(A) Transgenic mice overexpressing Pgam2 in a heart-specific manner were generated using the α-MHC promoter on a C57/BL6 background. The Pgam1 antibody (Abcam; Ab2220) detected both mouse PGAM1 and PGAM2 at similar sensitivities (Figure S1). Migration of the transgene product, the Pgam2 protein with a Myc-tag, was slower than that of the endogenous Pgam protein on a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel. Two of the 3 lines of Pgam2 mice (line 22 (L22) and line 38 (L38)) expressed different total Pgam protein levels in the heart (9.7-fold and 12.6-fold, when measurements included endogenous Pgam relative to levels in non-transgenic (NTg) mice, respectively). (B) The enzymatic activity of phosphoglycerate mutase was analyzed via an NADH-consuming spectrophotometric assay. The vertical axis indicated differences in optical density (O.D.) at 340 nm from the value of the negative control without heart tissue lysate. The half-time of ΔO.D. at 340 nm was 53% lower in Pgam2 mice than in NTg mice (n = 4 for each group).