Figure 2.
Rab11 regulates XK trafficking. (A) Colocalization of XK-EGFP with mCherry-Rab11 in striatal cells. SThdhQ7/Q7 cells were transfected with plasmids expressing XK-EGFP and with plasmids expressing dsRed-Rab4, dsRed-Rab5, dsRed-Vps35, or mCherry-Rab11. After treatment with β-cycloheximide, cells were processed for fluorescence microscopy. Shown are representative confocal images, which were captured individually for each channel and merged. Yellow structures indicate where the colocalization of XK-EGFP with mCherry-fused endosomal marker proteins took place. Scale bars: 10 μm (merge) and 2 μm (inset). (B) Pearson’s coefficient of colocalization. Digital images were analyzed with the JACoP plugin of the NIH ImageJ Fiji. Each symbol represents one cell. Data are mean ± SD. (C) Western blot of biotinylated proteins and postnuclear supernatants of SThdhQ7/Q7 cells transfected with plasmids expressing dNrab11 or dArab11 or empty vectors. Shown are blot analyses from one of the three individual experiments. (D) Densitometry for blot analyses in C. Data are mean ± SD. One-way ANOVA and post hoc Tukey’s analysis: F(2,6) = 44.26, P < 0.001; Tukey’s test * P < 0.01, # P < 0.001. (E) Effects of dominant active and inactive mutants of Rab11 on the subcellular distribution of XK-EGFP. STHdhQ7/Q7 cells were transfected with plasmids expressing XK-EGFP along with plasmids expressing mCherry-fused dArab11 or dNrab11. Confocal images showed that XK-EGFP and mCherry-dArab11 colocalized at small vesicular structures distributed throughout the cytoplasm and clustered at perinuclear regions, whereas the colocalization of XK-EGFP and mCherry-dNrab11 occurred at large tubulovesicular structures. Scale bars: 20 μm (merge) and 3 μm (inset). Source data are available for this figure: SourceData F2.

Rab11 regulates XK trafficking. (A) Colocalization of XK-EGFP with mCherry-Rab11 in striatal cells. SThdhQ7/Q7 cells were transfected with plasmids expressing XK-EGFP and with plasmids expressing dsRed-Rab4, dsRed-Rab5, dsRed-Vps35, or mCherry-Rab11. After treatment with β-cycloheximide, cells were processed for fluorescence microscopy. Shown are representative confocal images, which were captured individually for each channel and merged. Yellow structures indicate where the colocalization of XK-EGFP with mCherry-fused endosomal marker proteins took place. Scale bars: 10 μm (merge) and 2 μm (inset). (B) Pearson’s coefficient of colocalization. Digital images were analyzed with the JACoP plugin of the NIH ImageJ Fiji. Each symbol represents one cell. Data are mean ± SD. (C) Western blot of biotinylated proteins and postnuclear supernatants of SThdhQ7/Q7 cells transfected with plasmids expressing dNrab11 or dArab11 or empty vectors. Shown are blot analyses from one of the three individual experiments. (D) Densitometry for blot analyses in C. Data are mean ± SD. One-way ANOVA and post hoc Tukey’s analysis: F(2,6) = 44.26, P < 0.001; Tukey’s test * P < 0.01, # P < 0.001. (E) Effects of dominant active and inactive mutants of Rab11 on the subcellular distribution of XK-EGFP. STHdhQ7/Q7 cells were transfected with plasmids expressing XK-EGFP along with plasmids expressing mCherry-fused dArab11 or dNrab11. Confocal images showed that XK-EGFP and mCherry-dArab11 colocalized at small vesicular structures distributed throughout the cytoplasm and clustered at perinuclear regions, whereas the colocalization of XK-EGFP and mCherry-dNrab11 occurred at large tubulovesicular structures. Scale bars: 20 μm (merge) and 3 μm (inset). Source data are available for this figure: SourceData F2.

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