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. 2016 Jan 4;11(1):e0144561. doi: 10.1371/journal.pone.0144561

Fig 2. Generation and characterization of mice that overexpress rat sGC subunits in blood cells.

Fig 2

(A) Structure of the transgenes used to generate sGC transgenic mice. Human β-globin gene promoter (β-pro) is ligated with a mini β-LCR (open box), cDNA encoding either rat sGCα or sGCβ (black box), and an SV40 intron poly(A) signal sequence (SV40pA) (gray box). Black bar is equivalent to 1 kilo-base (kb). (B) Transgene expression in sGC transgenic mouse lines. Expression was examined by reverse transcriptase-PCR using 1 μg total RNA prepared from BM cells. PCR products of rat sGCα (R-sGCα), rat sGCβ (R-sGCβ), mouse sGCα (M-sGCα), and mouse sGCβ (M-sGCβ) are shown in (B)-(D). Lanes: 1, sGC-5; 2, sGC-7; 3, sGC-8; 4, sGC-9; 5, non-transgenic (Tg(-)) mice; M, molecular weight marker. In (B)-(D), mouse hypoxanthine phosphoribosyltransferase (HPRT) was used as an internal control for equal loading. Note: Mouse endogenous sGC mRNAs were undetectable in sGC-5 mice (Lane 1) by RT-PCR. (C) Transgene expression in various tissues of sGC-5 mice. Reverse transcriptase-PCR was performed using 1 μg total RNA prepared from various tissues; heart (lane 1); lung (2); liver (3); spleen (4); BM cells (5); peripheral RBCs (6); and peripheral leukocytes (7); M, molecular weight marker. (D) Developmental stage-specific expression of transgenes in sGC-7 mice and non-transgenic littermates. Reverse transcriptase-PCR was performed using 1 μg total RNA of yolk sac at 10.5 dpc (lanes 1&4); fetal liver at 14.5 dpc (2&5); and adult BM cells (3&6).