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. Author manuscript; available in PMC: 2014 Jul 30.
Published in final edited form as: Biochemistry. 2013 Jul 18;52(30):5103–5116. doi: 10.1021/bi4005914

FIGURE 8.

FIGURE 8

The effect of C-terminal truncation on the localization of three chimeric proteins, apically localized 50-64, basolaterally localized 50-80, and SLC13A3_15 with TLS. (A) Alignment of the C-termini of hSVCT1 and hSVCT2. The sequence within the light-grey box represents homology. The hSVCT1 residues under the black line were shown to be critical for apical localization of hSVCT1 (44). The hSVCT2 residues above the light-grey arrow were shown to be critical for membrane localization of both apical and basolateral SVCT proteins (4). The hSVCT1 residues beneath the darkgrey arrow were removed in the three constructs tested in this figure. (B) The TLS is visible for all three constructs in the confocal merged XY and YZ images. (C) The proportion of ascorbate uptake across the apical (Api) and basolateral (Baso) membrane of well-differentiated MDCK cells stably expressing each construct and grown on Transwells. (D) The efficiency with which each chimeric protein is incorporated into the cell membrane. (C,D) Data shown are means±SD of triplicate wells. Some error bars are too small to be visible.