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. 2017 Jan 3;292(7):2557–2570. doi: 10.1074/jbc.M116.768531

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© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Neu5Gc expression on polysialylated proteins. A, Neu5Gc incorporation into polySia. Delipidated brain homogenate from postnatal day 2–3 mice (littermates) was treated with EndoNF, and the released polySia-derived oligoSia fragments were hydrolyzed in acetic acid for DMB derivatization. Neu5Gc/Neu5Ac ratio in the obtained polySia-derived fractions (PolySia) or the delipidated homogenate before EndoNF treatment (Total) was analyzed by HPLC. Control, Cmah transgene-positive (no Cre) or Cre-positive (no Cmah transgene); Tg, NCmahTg. Each dot represents an individual animal, and bars indicate the means. B, expression of polysialylated NCAM. Brain lysate from postnatal day 1 mice were treated with EndoNF or PNGase F to cleave polySia or N-glycan, respectively, and then subjected to Western blotting with anti-NCAM (left) or anti-Neu5Ac polySia (right) antibodies. EGFP, Cmah transgene-positive (no Cre); Cre, Cre-positive (no Cmah transgene); Tg, NCmahTg; M, molecular weight marker; IB, immunoblot. Data are representative of two independent experiments that yielded similar results.