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The ASN4 glycosylation site is utilized in mammalian cells. Analysis of N-linked glycosylation of membrane-bound Ig5-FN1 expressed in mammalian cells. COS-1 cells were transfected with V5-tagged Ig5-FN1-TM-tail (Ig5-FN1) or mutant Ig5-FN1 with ASN5 and ASN6 or ASN4, ASN5, and ASN6 mutated to Gln (Ig5-FN1 Asn[5.6]Gln and Ig5-FN1 Asn[4.5.6]Gln, respectively). Cell lysates were subjected to PNGaseF digestion and analyzed by immunoblotting with anti-V5 epitope tag antibody.
