USF1 regulates ESC fate and hematopoietic differentiation by modulating bivalent domains of HS/PC-specific genes. DengChangwang LiYing LiangShermi CuiKairong SalzTal YangHui TangZhanyun G. GallagherPatrick QiuYi RoederRobert ZhaoKeji BungertJörg HuangSuming 2013 <p>(A) Western blotting assay of hSET1A levels in undifferentiated (day 0) and differentiated (day 13) ESCs comparing pcDNA vector transfected control and AUSF1 overexpressing cells. (B) FACS analysis of c-Kit and Tie2 expressing hemogenic endothelium cell population in pcDNA tranfected control and AUSF1 overexpressing ESCs upon hematopoietic differentiation at day 6. (C) FACS analysis of CD41 and c-Kit expressing early hematopoietic stem and progenitor population in pcDNA transfected control and AUSF1 overexpressing ESCs upon hematopoietic differentiation at day 10. (D) Real-time RT-qPCR analysis of hematopoietic markers and transcription factors, <i>c-Myb</i>, <i>CD150</i>, <i>CD34</i>, <i>Tal1</i>, <i>Runx1</i>, and <i>HoxB4</i> mRNA transcript levels upon hematopoietic differentiation comparing pcDNA transfected control and two AUSF1 overexpressing ES clones. Data are shown as mean ± SD. * P<0.05. (E) to (H). ChIP analyses of USF1 binding (E), <u>hSET1A</u> recruitment (F), H3K4me3 enrichment (G), and RNAPII loading (H) at the <i>HoxB4</i> promoter in AUSF1 overexpressing ES clones. Data are shown as mean ± SD. * P<0.05.</p>