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. 2010 Aug 30;285(45):35056–35067. doi: 10.1074/jbc.M110.170209

FIGURE 6.

FIGURE 6.

Insertion of specific NCAM FN1 sequences into the N-O chimera causes a switch from N-linked to O-linked polysialylation. A, COS-1 cells were transfected in duplicate with NCAM, N-O, N-O with NCAM acidic patch and α-helix sequences inserted, or N-O with acidic patch, α-helix, PYS, and QVQ insertions, along with ST8SiaIV-Myc. One sample was treated with PNGase F, and the other sample was left untreated. B, COS-1 cells were transfected in duplicate with N-O, N-O + PYS, and N-O + QVQ proteins along with ST8SiaIV-Myc. One sample was treated with PNGase F, and the other sample was left untreated. C, COS-1 cells were co-transfected with N-O or PYS mutants and ST8SiaIV-Myc. Upper panels, after immunoprecipitation of N-O proteins with anti-V5 epitope tag antibody, polysialylation was measured by immunoblotting with the anti-polysialic acid antibody, OL.28. Lower panels, relative N-O protein expression levels.