(A) Schematic representation of the strains used and crossed to obtain the endoglin specific KO mice in myeloid lineage. Luisa Ojeda-Fernández Lucía Recio-Poveda Mikel Aristorena Pedro Lastres Francisco J. Blanco Francisco Sanz-Rodríguez Eunate Gallardo-Vara Mateo de las Casas-Engel Ángel Corbí Helen M. Arthur Carmelo Bernabeu Luisa M. Botella 10.1371/journal.pgen.1005935.s001 https://plos.figshare.com/articles/figure/Mice_Lacking_Endoglin_in_Macrophages_Show_an_Impaired_Immune_Response/3139369 <p>The strain expressing Cre recombinase from the endogenous <i>Lyz2</i> locus (LysMCre) was crossed with the strain containing the floxed endoglin gene (<i>Eng</i><sup><i>fl/fl</i></sup>). Heterozygous mice for <i>Eng</i> floxed allele and positive for <i>Cre recombinase</i> were identified and crossed to obtain the three genotypes of interest. <b>(B)</b> Schematic representation of Cre recombinase action on endoglin floxed gene. Cre-mediated recombination results in deletion of the flanked sequence by LoxP sites in the myeloid cell lineage, including Mo, mature MΦ, and granulocytes. Cre action results in the deletion of exons 5–6 of endoglin gene. <b>(C)</b> Identification of mice genotypes by genomic PCR. Genomic PCR was performed with DNA from tails. The floxed endoglin allele (<i>Eng</i><sup><i>fl</i></sup>) was recognized by genomic PCR rendering a 566 bp product with primers Y and Z, and discriminated from the 411 bp product corresponding to the WT allele (<i>Eng</i><sup><i>wt</i></sup>) [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005935#pgen.1005935.ref051" target="_blank">51</a>]. The endoglin allele showing the exon 5–6 deletion (<i>Eng</i><sup>5-6</sup>) was detected by genomic PCR which gives rise to a 602 bp product using primers X and Y [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005935#pgen.1005935.ref051" target="_blank">51</a>]. <b>(D)</b> Efficiency of LysMCre-mediated lox P recombination in different tissues and PerC MΦ. PCR analysis of genomic DNA isolated from the indicated tissues of the three genotypes. The predicted amplicon sizes are indicated. The product of the amplification of <i>Eng</i><sup>Δ<i>5–6</i></sup> is undetectable in samples of <i>Eng</i><sup><i>wt/wt</i></sup><i>LysMCre</i> mice. PerC = peritoneal cavity.</p> <p>(TIF)</p> 2016-03-24 06:52:53 endoglin expression TGF Impaired Immune Response Endoglin Macrophages Show 57BL ENG vessel wall Phagocytic activity mice show vivo differentiation endoglin myeloid lineage endoglin gene Hereditary Hemorrhagic Telangiectasia role HHT 1 patients blood monocytes mouse line macrophage function ACVRL