Figure 4.
Defective trafficking of XK onto cell surfaces in HD cells. (A) Confocal images of SThdhQ7/Q7 and SThdhQ111/Q111 cells transiently transfected with plasmids expressing XK-pHluorin showed that pHluorin signals occurred at cell surfaces and intracellular compartments. Arrows point to cells with XK-pHluorin signals concentrated in the perinuclear regions. (B) Densitometry of pHluorin signals on the sharp edges of each of SThdhQ7/Q7 and SThdhQ111/Q111 cells showed a decline in the expression of XK-pHluorin on the surface of HD striatal cells. (C) Western blot analysis of biotinylated proteins and postnuclear supernatants of SThdhQ7/Q7 and SThdhQ111/Q111 cells with indicated antibodies. Shown are blot analyses from one of the three experiments. (D and E) Densitometry of signals of XK in postnuclear supernatants (total, D) and biotinylated XK (cell surfaces, E) for blot analyses in (C). (F) Images of XK staining in a brain section of WT and HD140Q/140Q mice. A series of consecutive coronal brain sections cut through the striatum were processed for labeling with antibodies for XK, with detergents omitted in all solutions to ensure the integrity of the plasma membrane. Images were captured from the nucleus accumubens of each brain section with the same settings. (G and H) Digital images from three brain sections for each of 3 WT and 3 HD mice were analyzed with the NIH ImageJ Fiji software to measure the cross-sectional area (G) and signal intensity (H) of striatal neurons immunoreactive to the XK antibody. Arrowheads in F indicate examples of neurons applied for densitometry and for measuring their cross-section areas. Scale bars: 20 μm (A), 100 μm (F), and 5 μm (inset in F). Data are mean ± SD. Each symbol in B, G, and H represents one cell and in D and E represents one experiment. Two-tailed Student’s t test. Source data are available for this figure: SourceData F4.

Defective trafficking of XK onto cell surfaces in HD cells. (A) Confocal images of SThdhQ7/Q7 and SThdhQ111/Q111 cells transiently transfected with plasmids expressing XK-pHluorin showed that pHluorin signals occurred at cell surfaces and intracellular compartments. Arrows point to cells with XK-pHluorin signals concentrated in the perinuclear regions. (B) Densitometry of pHluorin signals on the sharp edges of each of SThdhQ7/Q7 and SThdhQ111/Q111 cells showed a decline in the expression of XK-pHluorin on the surface of HD striatal cells. (C) Western blot analysis of biotinylated proteins and postnuclear supernatants of SThdhQ7/Q7 and SThdhQ111/Q111 cells with indicated antibodies. Shown are blot analyses from one of the three experiments. (D and E) Densitometry of signals of XK in postnuclear supernatants (total, D) and biotinylated XK (cell surfaces, E) for blot analyses in (C). (F) Images of XK staining in a brain section of WT and HD140Q/140Q mice. A series of consecutive coronal brain sections cut through the striatum were processed for labeling with antibodies for XK, with detergents omitted in all solutions to ensure the integrity of the plasma membrane. Images were captured from the nucleus accumubens of each brain section with the same settings. (G and H) Digital images from three brain sections for each of 3 WT and 3 HD mice were analyzed with the NIH ImageJ Fiji software to measure the cross-sectional area (G) and signal intensity (H) of striatal neurons immunoreactive to the XK antibody. Arrowheads in F indicate examples of neurons applied for densitometry and for measuring their cross-section areas. Scale bars: 20 μm (A), 100 μm (F), and 5 μm (inset in F). Data are mean ± SD. Each symbol in B, G, and H represents one cell and in D and E represents one experiment. Two-tailed Student’s t test. Source data are available for this figure: SourceData F4.

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