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Genetic and morphometric characterization of <i>C/X</i> mice.

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posted on 2015-09-15, 03:14 authored by Trevor L. Cameron, Katrina M. Bell, Irma L. Gresshoff, Lisa Sampurno, Lorna Mullan, Joerg Ermann, Laurie H. Glimcher, Raymond P. Boot-Handford, John F. Bateman
<p><i>(A)</i> RT-PCR on cDNA derived from femoral epiphyseal cartilage from wildtype (<i>Wt</i>) and <i>C/X</i> to detect the full-length form of <i>Xbp1</i> (<i>Xbp1</i><sub><i>FL</i></sub>) or the inactive form of <i>Xbp1</i>, lacking exon 2 (<i>Xbp1</i><sub><i>ΔEx2</i></sub>), and sequencing of cDNA from <i>C/X</i> femoral head cartilage to assay for the deletion of <i>Xbp1</i> exon 2. <i>(B)</i> PCR for residual <i>loxP</i> site downstream of the p.Asn617Lys <i>Col10a1</i> coding sequence using genomic DNA derived from <i>Wt</i> and <i>C/X</i>. <i>(C)</i> Alizarin red S/Alcian blue staining of skeletal preparations from newborn, 1 week, and 2 week wildtype (<i>Wt</i>), <i>Xbp1</i><sup><i>CartΔEx2</i></sup>, <i>ColX</i><sup><i>N617K</i></sup> and <i>C/X</i> mice. <i>(D-F)</i> Quantification of <i>(D)</i> femoral and <i>(E)</i> tibial length, and <i>(F)</i> intercanthal distance (ICD) from legs from 2 week <i>Wt</i> and mutant mice–<i>Wt</i>, N = 8; <i>Xbp1</i><sup><i>CartΔEx2</i></sup>, N = 8, <i>ColX</i><sup><i>N617K</i></sup>, N = 6; <i>C/X</i>, N = 8; statistical analysis performed using Student’s <i>t</i> test.</p>

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