Characterization of the lactational defect in <i>Ndst1</i>-deficient mammary glands.

<p>Quantitative RT-PCR and Western blotting were used to characterize lactational capacity of <i>Ndst1</i>^f/f<i>MMTVCre</i>⁺ females. (<b>A</b>) RNA isolated from d1L <i>Ndst1</i>^f/f<i>MMTVCre</i>^– and <i>Ndst1</i>^f/f<i>MMTVCre</i>⁺ mammary glands was analyzed by quantitative RT-PCR for the expression of whey acidic protein (WAP), β-casein, and keratin 18, genes specifically expressed by mammary epithelial cells. Data was normalized to the expression of GAPDH transcripts present in the sample. (<b>B</b>) Western blotting with antibodies to keratin 7 in d1L glands further confirmed that the <i>Ndst1</i>^f/f<i>MMTVCre</i>⁺ mammary glands have a reduced population of epithelia. (<b>C</b>) Western blotting with antibodies to mouse milk from d1L glands shows that milk production was diminished in <i>Ndst1</i>^f/f<i>MMTVCre</i>⁺ glands. Blotting of d14P extracts showed that the antibodies were selective for milk protein except for a minor band at 67 kDa.