Figure 2. Inversion of the Notch3^COIN allele in the germ line causes osteopenia.

A. DNA was extracted from tail of heterozygous Notch3^em2.1Ecan (Hprt-Cre;Notch3^INV/WT) and wild type littermate controls and Notch3^COIN inversion was documented by gel electrophoresis of PCR products obtained with primers specific for the Notch3^INV allele. Arrows indicate the position of the 495 base pair (bp) amplicon. B. Total RNA was extracted from calvariae of 1 month old Notch3^em2.1Ecan mutants (closed circles) and wild type littermate controls (open circles), and expression of the Notch3 ^PEST and Notch3^ΔPEST mRNA measured by qRT-PCR. Transcript levels are reported as copy number corrected for Rpl38 mRNA levels. Bars represent means and ranges S.D.; n = 9 for control, n = 11 for Notch3^em2.1Ecan, all biological replicates. C. Weight in gm and femoral length in mm of 1 month old Notch3^em2.1Ecan (closed circles) and wild type control littermates (open circles). Bars represent means and ranges SD; n = 5 for control males and females, n = 6 for Notch3^em2.1Ecan males and females. D. Representative μCT images of femoral proximal trabecular bone and midshaft cortical bone of 1 month old control and Notch3^em2.1Ecan mice. The scale bar in the right corner represents 500 μm. Complete data set in Table 3. *Significantly different between control and Notch3^em2.1Ecan, p < 0.05 by unpaired t-test for Panel B and ANOVA for Panel C.