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. 2016 Aug 12;36(17):2282–2299. doi: 10.1128/MCB.01077-15

FIG 4.

FIG 4

Generation and characterization of fro/fro; Acan-Smpd3 mice. (A) Schematic representation of the Acan-Smpd3 transgene construct. Arrows, positions of the primers used for genotyping; β, β-globin intron; pA, SV40 polyadenylation signal sequence. (B) A 610-bp amplicon was obtained from the PCR analysis of the transgenic DNA. (C) (Top) Semiquantitative RT-PCR analysis confirming the cartilage-specific expression of the transgene (TG). (Bottom) Hprt expression is shown as a loading control for cDNA. (D) Skeletal preparations of WT, fro/fro, and fro/fro; Acan-Smpd3 newborn pups stained with alizarin red and alcian blue show that in fro/fro; Acan-Smpd3 mice there is a partial correction of the skeletal deformities seen in fro/fro mice. Arrows, limb deformities. (E) Magnified view of the forelimbs and hind limbs of skeletal preparations of newborn WT, fro/fro, and fro/fro; Acan-Smpd3 pups. (F) X-ray analysis of the femur and tibia of 1-month-old mice showing that the long bones of fro/fro; Acan-Smpd3 mice are significantly shorter in length and slightly bent (arrow) compared to the control bones. **, P < 0.01. (G) Vertebral sections of 1-month-old mice stained with VK-VG showing a higher osteoid (unmineralized bone) volume over bone volume (OV/BV) in fro/fro; Acan-Smpd3 mice than the control mice.