Impact of Hybrid and Complex N-Glycans on Cell Surface Targeting of the Endogenous Chloride Cotransporter Slc12a2

Figure 1

NKCC1a transcripts are expressed in COS7 cells. (a) Representation of the last nine exons (boxes) of the human SLC12A2 gene. Alternative spliced exon 21 is indicated as a grey box in NKCC1a (NM_001046). Introns are represented with V-shaped lines. White boxes represent 3′-UTRs. Underneath, shown are the primer sets used to coamplify NKCC1a and NKCC1b mRNAs (represented as opposite arrowheads connected by a line). These primers are labeled according to the size in bp of the expected amplicons (see text). (b) Shown is a representative 2% agarose gel where bands of predicted sizes for NKCC1a are detected (608 bp and 516 bp). (c) In human brain, bands of predicted sizes for NKCC1a and NKCC1b are coamplified: 608 bp + 560 bp and 516 bp + 468 bp, respectively. As positive control, RT-PCR reactions amplified GAPDH (555 bp). As negative control, RT-PCR reactions were performed using H₂O instead of total RNA and GAPDH-555 primer sets. (d) Representative DNA sequence chromatogram corresponding to amplicons obtained with primer set NKCC1-608a/560b encompassing exons 20, 21, and 22 in NKCC1 mRNAs. The sequences obtained from COS7 cells were aligned against chimpanzee (XM_526998) and human NKCC1 transcripts of reference. The sequence difference found in that region is highlighted.

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