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

<p><b>(A)</b> 8-bromo-cGMP induces human γ-globin mRNA expression in human BM progenitors. P value: *, <i>P</i><0.05 compared with control. <b>(B)</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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144561#pone.0144561.s004" target="_blank">S1 Text</a>. Representative elution profiles are shown from 3 independent experiments that gave similar results. <b>(C) (D)</b> 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. <b>(E)</b> Summary of human β-like globin expression in YAC mice and sGC-YAC mice during development. P value: *, <i>P</i><0.05 compared with YAC mice. <b>(F)</b> 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 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144561#pone.0144561.ref031" target="_blank">31</a>] 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. <b>(G)</b> 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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144561#pone.0144561.s004" target="_blank">S1 Text</a>. 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. *, <i>P</i><0.05.