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

Fig 6. Examination of human β-like globin expression in sGC transgenic mice carrying a YAC clone (sGC-YAC) including the human β-globin gene locus.

Fig 6

(A) 8-bromo-cGMP induces human γ-globin mRNA expression in human BM progenitors. P value: *, P<0.05 compared with control. (B) Expression of human γ-globin expression in the fetal liver (14.5 dpc) of YAC mice and sGC-YAC mice. Human β-globin was used as the internal control to compare the γ-globin expression levels of YAC and sGC-YAC mice. Globin chain analysis in fetal livers was performed by reverse-phase HPLC as described in S1 Text. Representative elution profiles are shown from 3 independent experiments that gave similar results. (C) (D) Expression of human γ-globin expression in adult YAC mice (C) and sGC-YAC mice (D). Globin chains in peripheral blood prepared from YAC or sGC-YAC mice were analyzed by reverse-phase HPLC. In (D) upward bold arrow indicates the position of Gγ-globin in sGC-YAC mice. Note: Gγ-globin, but not Aγ-globin, was detected in RBCs from adult sGC-YAC mice. (E) Summary of human β-like globin expression in YAC mice and sGC-YAC mice during development. P value: *, P<0.05 compared with YAC mice. (F) Detection of human γ-globin in mouse RBCs. Total cell lysates (10 μg) prepared from mouse RBCs were analyzed in 12% SDS-PAGE gels by immunoblotting as described [31] Lanes: 1, RBCs from fetal livers of YAC mice (14.5 dpc); 2, peripheral blood RBCs from adult YAC mice; 3, peripheral blood RBCs from adult sGC-YAC mice. (G) Recruitment of RNA polymerase II to β-like globin gene promoters in sGC-YAC mice. ChIP assays were performed using spleen-derived erythroblasts as described in S1 Text. Upper panel: 1, input DNA; 2, non-specific IgG antibody; 3, brain; 4, sGC mice; 5, sGC-YAC mice; 6, YAC mice. Lower panel: 1, input DNA; 2, non-specific IgG antibody; 3, brain; 4, sGC mice; 5, fetal liver cells (14.5 dpc) of sGC-YAC mice; 6, YAC mice;7, adult erythroid cells from sGC-YAC mouse. Representative results are shown from experiments that were repeated at least 3 times. *, P<0.05.