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. 2020 Jul 28;39(16):e103009. doi: 10.15252/embj.2019103009

Figure 3. Rab11a labels a distinct subset of multivesicular bodies and their intraluminal vesicles in HCT116 colorectal cancer cells.

Figure 3

  • A–D
    Confocal images of fixed HCT116 cells, with boxed regions enlarged to the right. DAPI (grey) marks nucleus. Rab11a antibody is isoform‐specific except in (F). (A) Rab11a (yellow) is located in compartments distinct from the late endosomal and lysosomal marker, LAMP2 (magenta). (B) CD63 (magenta) predominantly co‐localises with the late endosomal and lysosomal marker, LAMP1 (yellow). (C) Rab11a (yellow) is located in compartments distinct from CD63 (magenta) under glutamine‐replete conditions. (D) Rab11a (yellow) is located in compartments distinct from CD63 (magenta) under glutamine‐depleted conditions.
  • E
    Super‐resolution 3D‐SIM image of fixed HCT116 cell expressing GFP‐Rab11a (yellow), and stained with CD63 (magenta). DAPI (grey) marks nucleus. Boxed Rab11a‐positive compartments, which frequently cluster, are magnified in Merge Zoom. This panel is further magnified in Merge Zoom × 2, revealing GFP‐Rab11a (arrows in right panel) inside compartments.
  • F
    Wide‐field fluorescence image of fixed HCT116 cells, stained with Rab11a (yellow) and Rab7 (magenta), expressing constitutively active GFP‐tagged Rab5 (GFP‐Rab5CA; cyan), which stalls endosomal maturation and produces enlarged Rab5‐positive endosomes. One of these is boxed in the Merge and magnified in Merge Zoom, revealing internal puncta marked by Rab11a (arrows) and Rab7 (arrowheads) and limiting membrane subdomains of Rab11a (yellow arrowhead) and Rab7 (magenta arrowhead). DAPI (grey) marks nuclei.
Data information: Scale bars in A–F (5 μm), in Merge Zoom (1 μm) and in Merge Zoom × 2 (0.5 μm).