Figure 5.
Figure 5. Subcellular localization of predicted new CCV, retromer, and AP-4–associated proteins. The proteins shown in Fig. 4 were localized by immunofluorescence microscopy. In the case of REPS1, the endogenous protein was detected with a specific antibody. Other proteins were myc- or GFP-tagged as indicated, and transiently expressed. Colocalization with intracellular or plasma membrane clathrin-coated structures was investigated by double labeling with antibodies against AP-1 γ or AP-2 α, respectively. C10orf88-myc colocalizes with AP-1 and AP-2, which necessitated imaging in two different focal plains. For reasons of antibody compatibility, CALM was used in this case to define clathrin-coated structures at the plasma membrane. Overlays show new proteins in red and reference proteins in green. Bars: (large panels) 10 µm; (insets) 2 µm.

Subcellular localization of predicted new CCV, retromer, and AP-4–associated proteins. The proteins shown in Fig. 4 were localized by immunofluorescence microscopy. In the case of REPS1, the endogenous protein was detected with a specific antibody. Other proteins were myc- or GFP-tagged as indicated, and transiently expressed. Colocalization with intracellular or plasma membrane clathrin-coated structures was investigated by double labeling with antibodies against AP-1 γ or AP-2 α, respectively. C10orf88-myc colocalizes with AP-1 and AP-2, which necessitated imaging in two different focal plains. For reasons of antibody compatibility, CALM was used in this case to define clathrin-coated structures at the plasma membrane. Overlays show new proteins in red and reference proteins in green. Bars: (large panels) 10 µm; (insets) 2 µm.

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