<i>Jpx</i> transgenes induce <i>Xist</i> expression in mESCs using both <i>trans</i> and <i>cis</i> mechanisms. Sarah Carmona Benjamin Lin Tristan Chou Katti Arroyo Sha Sun 10.1371/journal.pgen.1007378.g001 https://plos.figshare.com/articles/figure/_i_Jpx_i_transgenes_induce_i_Xist_i_expression_in_mESCs_using_both_i_trans_i_and_i_cis_i_mechanisms_/6228752 <p>(A) Map of the X-inactivation center (Xic) with 90kb Tg(Jpx) and 120kb Tg(Jpx, Xist) transgenes and the probes used for Fluorescence <i>in situ</i> Hybridization (FISH). (B) Combined RNA-DNA FISH for <i>Xist</i> (green, FITC) and <i>Xpct</i> (red, Cy3) on female mESC at differentiation days 0, 2, and 4. Top: Control female ESCs transfected with Tg(pSKYneo+), a plasmid that does not contain X-chromosome sequence but provides the same neomycin resistance as Tg(Jpx, Xist); bottom: Tg(Jpx, Xist) transgenic female mESC line #7, which has a single-copy Tg(Jpx, Xist) integrated in an autosome; right: schematic of <i>Xist</i> expression at Day 4. Green wavy line, <i>Xist</i> RNA; Red dot, <i>Xpct</i> (X-Chr.) DNA locus; Open arrowhead: Tg(Jpx, Xist) transgenic site. Scale bar: 2μm. (C) <i>Xist</i> cloud frequency throughout differentiation. **, <i>P</i><0.01; ***, <i>P</i><0.001, from a chi-square test comparing total <i>Xist</i> cloud counts in wildtype and transgenic cells at each differentiation day. Sample sizes and additional Tg(Jpx, Xist) mESC lines are included in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007378#pgen.1007378.s001" target="_blank">S1 Fig</a>.</p> 2018-05-07 17:38:26 Xist expression competence factor cis mechanisms Mammalian X chromosome dosage compensation balances Jpx copy number RNA