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. 2013 Apr 4;8(4):e61112. doi: 10.1371/journal.pone.0061112

Figure 6. N-Glycolsylation deficiency of the KCC3-E289G mutant.

Figure 6

A) Western blot analyses of KCC3-E289G mutant in HEK 293FT cells and Xenopus laevis oocytes compared to wild-type KCC3 in HEK 293FT cells, wild-type and mutant CHO cells, and Xenopus laevis oocytes, using rabbit polyclonal anti-KCC3 antibody. CHO-Lec1 cells have mutation in N-acetylglucosaminyl transferase, whereas CHO-Lec8 and CHO-Lec2 have deficient galactose and sialic acid transporters, respectively. Two independent experiments are shown. Experiment was performed 4 times. B) Scheme represents the main steps in N-linked oligosaccharide biosynthetic pathway. First, core Glc-Nac-Glc-Nac-Man with branched mannose residues are added to the Asparagines in the ER. In the Golgi, mannose molecules are replaced by acethylglucosamyl groups, followed by the addition of galactose and sialic acid groups. These steps require the availability of galactose and sialic acid in the cells, which is prevented in the mutant CHO cells by elimination of specific transporters.