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. 2013 Aug 28;288(43):31080–31092. doi: 10.1074/jbc.M113.469189

FIGURE 3.

FIGURE 3.

Expression and transport function of four FBS-associated mutants of hGLUT2. A, expression of hGLUT2 wild type (WT) and four FBS-associated mutants (G20D, S242R, P417L, and W444R) in mhAT3F cells. Cells transfected with a pCMV-hGLUT2-HA-IRES-hrGFP construct are identified by the expression of the GFP. Subcellular location of hGLUT2 (red in the left panels and white in the right panels) is revealed with an antibody to HA epitope in permeabilized cells. Nuclei are stained with DAPI (blue). Note that G20D hGLUT2 cannot be detected, and S242R hGLUT2 is not targeted at the plasma membrane. Scale bar corresponds to 25 μm. B, Western blot analysis of membrane fractions from mhAT3F cell transfected or not transfected (NT) with different hGLUT2 constructs. hGLUT2 expression is revealed with an antibody to HA epitope. E-cadherin is used as a loading control of membrane fractions. Note that G20D or S242R hGLUT2 cannot be detected in membrane fractions. C, membrane expression of WT, S242R, P417L, and W444R hGLUT2 in Xenopus oocytes injected with the corresponding cRNAs. D, noncorrected uptake of 2-DOG by Xenopus oocytes injected with water or injected with WT, S242R, P417L, or W444R hGLUT2 cRNA.